Antimicrobial compositions and methods of making the same

ABSTRACT

The present disclosure comprises antimicrobial compositions and devices comprising silver compounds that resist heat and light discoloration. In one aspect, the said compounds comprise silver and at least one s-triazine ring or moiety. In another aspect, the antimicrobial compositions are hydrogels that are effective against broad spectrum of common pathogens including MRSA and VRE and are suitable for treating human or animal wounds and burns. The methods of the present disclosure comprise treating medical and non-medical devices and articles with compositions comprising the silver compounds to impart antimicrobial property.

RELATED APPLICATION

This application claims priority from U.S. provisional patentapplication Nos. 61/854,850 filed on 2 May 2013 and 61/967,002 filed on8 Mar. 2014, respectively.

FIELD

The present disclosure broadly pertains to antimicrobial compositionsand devices comprising silver and specifically compositions for treatingwounds and burns and the methods for making them.

BACKGROUND AND SUMMARY

To promote faster healing of infected wounds and burns, reduction ofbio-burden is the first step. In quantitative terms, the bio-burden ofinfected wounds can reach as high as a million colony forming units(cfu) per gram of tissue. Therefore, rapid disinfection of the woundsfollowed by maintenance of low bio-burden is particularly attractive.

The most widely accepted clinical practice for reducing the bio-burdenof the wounds is to cover them with dressings infused with antimicrobialcompounds. The choice of wound dressing is dependent on the state of thewound; for badly infected wounds the dressings that releaseantimicrobial actives to the wounds very rapidly are preferred to causebacterial count to drop to negligible levels. Thereafter, with optimalmoisture management the body's immune system takes over to acceleratehealing. A commercial product, Acticoat® is one such dressing thatrapidly releases a lethal bolus of ionic silver. While such burst ofionic silver kills bacteria, it also stains skin and in the short termoften retards wound healing.

Alternately, there are products that deliver the antimicrobial activesto the wound site more slowly over time. In such case, the bio-burdendecrease takes place slowly but without interference with the body'snatural healing process. Additionally, these products are compoundedwith agents that aid healing and provide for moisture management.Examples of such products include antimicrobial hydrogels (SilvaSorb®,SilverSept®, Normlgel Ag® and Elta®) and antimicrobial sheet dressings(SilvaSorb® and Covalon®)

However, in these products, especially the hydrogels, the amount ofactive silver compound is kept low so that in topical use they are nottoxic to the skin cells. In hydrogels, due to their complex compositionsand high viscosities, lower amounts of the silver compound often leadsto uneven performance because the active species (Ag⁺ ions) is sometimesprematurely reduced to elemental silver (Ag⁰) which is inactive againstmicroorganisms. The premature reduction often occurs in the hydrogelcompositions in packaged form during storage due to various factors suchas interaction with packaging material and changes in environmentconditions. Thus, the silver containing compositions currently on themarket perform differently when made fresh and may fail as they approachthe end of their shelf life. To compensate for decreased activity of thehydrogel product nearing its end of useful life, the formulators oftenincrease the amount of active ionic silver. However, increased amountsof ionic silver in the hydrogels increases the risk of prematurereduction due to various factors mentioned earlier. The reduction of theactive silver compound to inactive elemental silver in hydrogelcompositions is accompanied by undesirable discoloration. In somesituations, the hydrogels undergo a change in gel pH to acidic resultingin increased stinging and irritation to the patient's skin. Thisparticularly is extremely undesirable to persons with sensitive skin orthose with burns.

While some silver containing antimicrobial products such as Acticoat®dressings are dark colored and have been acceptable, consumerpreferences do not permit dark colored or discolored hydrogels. Thus,antimicrobial compositions, particularly hydrogels that carry greateramounts of active silver compound(s) and yet are not dark colored whenmade or darkened prematurely in the packaged form may be useful.Furthermore, antimicrobial compositions that possess pH near neutral andare robust against pH drift into the acidic range may also be useful. Inaddition, antimicrobial hydrogel compositions that are clear to aid inthe monitoring of healing wounds and that are able to provide moisturemanagement may provide further utility in practice.

To provide a robust and effective antimicrobial hydrogel compositionstarts with a robust and effective antimicrobial active agent. Among theactives, in theory silver is quite effective because at therapeutic uselevels it is non-toxic and there is history of its safe use amongclinicians for over hundred years. Besides, there is practically no riskof common pathogens developing resistance to silver due to itsmulti-prong disruption of the bacterial growth cycle. In contrast, thepopular antibiotics are already becoming ineffective as resistantstrains of microorganisms are slowly emerging, which is an unintendedoutcome of their overuse. Other antimicrobials such as biguanides andchlorohexidine compounds may be potentially useful but they havetoxicity issues and so may not work well.

However, despite the promise of silver, products with silver have notbeen as widespread in use. That's because an overwhelming majority ofsilver compounds are prone to heat and light induced discoloration andhence are not robust. Often those that are sufficiently resistant aresparingly soluble in water, e.g., silver sulfadiazine. For example,since the introduction of silver sulfadiazine forty years ago, therehave been no reports of any silver chemistry that have matched orexceeded its discoloration resistance. Because of poor solubility inpractically all solvents, silver sulfadiazine has been met with limitedsuccess. Given that there have been reports of silver sulfadiazine asnot being as effective against microorganisms that have developedresistance to sulfonamides, going forward it is less likely to be theactive silver compound of choice for device manufacturers andformulators. Further, the solubility problem in general can lead toproduct quality issues, which may increase use levels to achieveefficacy and therefore make manufacturing tricky. While there is nomatch to silver's broad spectrum efficacy, silver containing productswhen in contact with body parts or skin can cause staining Finally, theunpredictability of discoloration in silver containing devices may leadto poor yields in manufacturing, quality issues and a short shelf life.Various approaches to stabilization of silver in devices andcompositions have been developed, but they have had limited utility dueto their device specificity and limited implementation. Thus, anantimicrobial silver compound or a group of compounds that can providemore broadly robust resistance to heat and light induced discolorationand yet be relatively straightforward to incorporate into devices andcompositions including hydrogels is lacking.

The inventor has recognized these issues and herein describesantimicrobial compositions that comprise silver cyanurate derivativesthat hitherto were not investigated as antimicrobial actives.Antimicrobial devices comprising said compounds are also contemplated bythe present disclosure. In one example, the antimicrobial compositionsare hydrogels. The hydrogels are smooth, viscous, thixotropic, clear totranslucent, readily spreadable under shear forces generated in topicaluse. Features of said antimicrobial compositions are clarity, ability toresist light and heat induced discoloration despite comprising activesilver compounds at higher loadings. Some example hydrogel embodimentsof the present disclosure are able to resist discoloration due tosunlight exposure or elevated temperatures of steam sterilizationwithout compromising antimicrobial activity. The ability of saidhydrogel compositions to withstand sunlight and elevated temperatureswithout discoloration while maintaining its antimicrobial effectivenessis a distinguishing feature of the present disclosure. Put another way,the robust thermal stability of said hydrogels precludes special storageconditions or shipping requirements and translates into practically anindefinite shelf life.

The antimicrobial hydrogel compositions of the present disclosure arenon-staining to the skin and at use levels envisioned non-toxic tohumans and animals. They possess effective broad spectrum antimicrobialactivity against substantially all common pathogens: bacteria includingMRSA and VRE, yeasts and fungi, but at higher silver loadings may beeffective against, amoeba, protozoa, virus, etc. The said hydrogelcompositions are suitable for use in the treatment of acute and chronicwounds that diabetics suffer, first and second degree burns and woundson mucous membranes. When compounded appropriately they are effectiveand safe OTC products to treat minor cuts, burns and abrasions withminimal risk of staining. The hydrogels promote and accelerate woundhealing by reducing bio-burden and promoting moisture management of lowto moderate exuding wounds.

The present disclosure further provides methods of using a group ofsilver cyanurate compounds as antimicrobials. The compounds are inert toheat (steam sterilization temperatures) and light (direct sunlight),relatively easy to synthesize and incorporate into compositions anddevices. To impart antimicrobial properties to compositions, they arederived simply by reacting metal cyanurates with soluble silver salts insolutions in situ or formed separately either individually or as amixture and then compounded. Though sparingly soluble in water, whencompounded into antimicrobial hydrogels at effective use levels,surprisingly they do not adversely affect gel transparency. Methods ofmaking antimicrobial devices and compositions comprising said compoundsfor use as wound care products or patient care products are contemplatedby the present disclosure. Examples of non-medical devices and theirapplications are provided.

DETAILED DESCRIPTION

The above advantages and other advantages, and features of the presentdescription will be readily apparent from the following DetailedDescription. It should be understood that the summary above is providedto introduce in simplified form a selection of concepts that are furtherdescribed in the detailed description. It is not meant to identify keyor essential features of the claimed subject matter, the scope of whichis defined uniquely by the claims that follow the detailed description.Furthermore, the claimed subject matter is not limited toimplementations that solve any disadvantages noted above or in any partof this disclosure.

Antimicrobial compositions that comprise silver are contemplated. Thecompositions are amorphous topical formulations that are suitablyeffective as broad spectrum antimicrobials against substantially allcommon pathogens: bacteria (gram positive and gram negative), anaerobes,yeasts and fungi. Though said compositions are contemplated for topicaluse, their use on mucous membranes in humans and animals, e.g., buccalcavity, is within the scope of the present disclosure. Depending on theamount of the antimicrobial active silver compounds in the compositions,they may also be effective against other organisms such as viruses,amoeba, protozoa, etc.

Antimicrobial Hydrogel Compositions

In one example, the compositions of the present invention are hydrogels,though other forms ranging from dilute suspensions (very low viscosity)or solutions to concentrates or dough-like compositions (extremely highviscosity) are not outside the scope of the present disclosure. Ashydrogels, they are amorphous and therefore intuitive to use in anytopical application. In one aspect, the compositions are thixotropicgels and possess yield stress. Such gels exhibit a rapid decrease inapparent viscosity when disturbed by stirring or even vigorous shaking.However, when the disturbance is removed, the apparent viscosity isrestored and maintained during the dormancy state. The yield stress isthe value that is overcome for transition from the gel (structuredstate) to sol (the flow or unstructured state) and dictates the easewith which the gel compositions can be spread. In one aspect, thehydrogel compositions of the present disclosure possess yield stressthat range between 0 and 1000 Pa at 20-25 C, more preferably between 5and 750 Pa and most preferably between 50 and 400 Pa. In addition, lowyield stress permits ease of dispensing from packaged tubing. Asdescribed herein, the clear topical hydrogel may comprise anantimicrobial compound comprising silver and an s-triazine ring. In oneexample, the clear topical hydrogel may comprise hydrogen peroxide inaddition to the silver cyanurate active agent. In some topicalapplications, the hydrogel is a thixotropic hydrogel and may have ayield stress in a range of 0 and 1000 Pa.

Another attractive property of the hydrogel compositions is thetransparency. This attribute is desired by clinicians as it allows themto monitor from outside (through a thin film moisture permeabledressing) the progress of wound healing without having to resort topainful (to the patient) dressing changes to examine wound shrinkage.Interestingly, the transparency in the hydrogels is maintained despitethe presence of active silver compounds to levels as high as 1000 ppm.Considering that the silver actives of the present disclosure aresparingly soluble in water the transparency observed in said hydrogelsis quite remarkable.

The hydrogels of the present disclosure have low to moderate absorptioncharacteristic that helps with moisture management of low to moderateexuding wounds. In heavily exuding wounds, excess moisture can oftenretard the healing process. Certain commercial products possess muchhigher absorption capacity compared to the hydrogels of the presentdisclosure. One embodiment of the present disclosure overcomes thelimitation of moderate water absorption capacity of the presentantimicrobial hydrogels. The embodiment utilizes a kit comprising theantimicrobial hydrogel (which can be sterile) and a sterile high watercapacity bandage, e.g., a hydrophilic gel sheet or foam that incombination is used to effectively treat heavily exuding wounds. Thehigh absorption capacity bandage helps maintain moisture in the woundwhile the antimicrobial silver hydrogel ensures a bacteria freeenvironment, thus accelerating wound healing.

Certain wound hydrogels on the market contain microparticles ofcross-linked hydrophilic polymer to enhance water absorption capacity.However, they are grainy to feel and often leave gel residue on theskin. In contrast, the present hydrogels are smooth to feel and can berubbed into the skin akin to vanishing creams. This feature is quiterelevant for OTC products meant to treat minor cuts, burns or abrasionsbecause the presence of product residue is not desired by young patientswho are end users.

Unlike some antimicrobial hydrogels on the market, the antimicrobialhydrogels of the present disclosure possess pH near neutral (7-8 range)and maintain their pH reasonably stable. They typically are free fromthe problem associated with stinging of skin caused by acidic pH. Ifincorrectly formulated, hydrogel products can cause pH drift into acidic5-6 range over time. To mitigate the risk associated with stingingcaused by acidic pH from ever happening with the antimicrobial hydrogelsof the present disclosure, adding a painkilling compound, e.g., from thebenzocaine family is within the scope of the present disclosure. Butdepending on the application, the hydrogel compositions may possess pHbetween 2 and 10. Such variations are also contemplated by the inventor.For instance, the compositions with acidic pH (5 or less) may bepreferred in the treatment of fungal infection of nails.

For any topically applied antimicrobial product over the breached skin,the toxicity is of concern. Repeated use can cause the active compoundto enter the body and do unintended harm. In the wound environment, theactive levels can interfere with biochemical processes associated withhealing. The antimicrobial hydrogels of the present disclosureincorporate silver compounds as active agents that are relatively safeat the levels contemplated for a wound care product intended for 3-5days of use. In addition, the active silver compounds comprise moietiesthat have been used in commerce and have an acceptable safety record.For instance, the LD₅₀ value for rats in the case of cyanuric acid is7700 mg/kg body weight (BW). In contrast, several common chemicalsencountered in daily life such as boric acid (medicinal uses), benzoicacid or sodium benzoate (preservative) and salicylic acid (medicinaluses) have LD₅₀ (rats) values lower than that for cyanuric acid meaningthey are more toxic. Further, the use of hydrogel compositions isprimarily topical. Moreover, the compositions are viscous gels thatoffer a greater resistance to the diffusion of cyanurate anions orsilver cations for entry into the body through breached skin. In thisway, as one example, the amount of cyanuric moiety entering the body maybe a thousand fold less than the toxicity threshold. As such, underextreme scenarios such as a condition where a burn victim has 40% bodycoverage with topical gel application that lasts over several days, thesilver cyanurate derivatives may be considered practically non-toxic,even in such use levels contemplated by the present disclosure. As oneexample, identified use levels correspond to silver element equivalenceof 2000 ppm or less with additional use levels being less than 1000 ppm.As another example, an amount of silver in the hydrogel may be between50 and 1000 ppm based on a weight of the hydrogel.

The antimicrobial hydrogel compositions are suitable whenever there issufficient need to provide antimicrobial conditions to sustain bacteriafree environments. In a broader sense, the antimicrobial compositionsprovide an added layer of protection against bacterial or fungalcontamination. They are especially useful in the treatment of (a)chronic and acute wounds and (b) first and second degree burns. Forexample, chronic wounds are characterized by a prolonged period ofinflammation and delays in wound healing and repair, which often resultsfrom contamination by microorganisms. In contrast, heavily infectedwounds have very high bio-burden with bacterial counts in excess of onehundred thousand cfu per gram of tissue. In both cases, theantimicrobial compositions provide therapeutic action by effectivelyeliminating or lowering bio-burden rapidly so that the body's naturalhealing processes can be initiated.

Because of the low to moderate thixotropic yield stress of theantimicrobial hydrogel compositions, they can be more easily applied asa thin layer under shear forces generated by fingers or hand. Thus, theyare suitable for use to prevent infections on fresh burn skin areascharacterized as 1^(st) and 2^(nd) degree burns without the risk ofcausing excruciating pain to the subject. When compounded to lowesteffective silver levels in said compositions they can be used to treatminor cuts, abrasions and burns. As an added benefit, the hydrogelcompositions do not cause skin stain even when applied areas are exposedin the daylight. This aspect can be valuable in an OTC product becausekids often suffer such injuries playing outside and applying thecompositions over injured areas would not preclude play activity becauseof fear of skin staining. In hospital settings, they are suitable to usewithout the risk of staining of patient garments, bed sheets andmattress coverings.

The medical applications of the antimicrobial hydrogel compositionscomprising silver extend beyond wound healing. For example, theantimicrobial formulations with pH (5 or less) may be useful in thetreatment of fungal infections of the nail and nail bed (onychomycosis)by eliminating causative dermatophytes. Alternately, embodiments of thepresent disclosure may be used to treat dermal conditions such as acne,rosacea, jock itch, and athlete's foot caused by the anaerobic bacteria(acne), demodex mites (rosacea), and fungi (jock itch and athlete'sfoot). Another embodiment of the antimicrobial compositions is as anultrasound gel used in pregnancy monitoring. The compositions can renderthe applied skin area bacteria free during checkup and can havesufficient sound transmittivity due to the use of Laponite® syntheticclay as thickener. They can adequately replace current ultrasound gelproducts as they are also non-staining.

Besides hydrogels, other forms of amorphous compositions comprisingactive silver compounds are contemplated by the inventor. Suitableexamples are suspensions, solutions, bioadhesive or adhesivecompositions (U.S. Pat. No. 4,914,173), polymer solutions, lotions,creams, o/w or w/o emulsions, emulgels, salves, pessaries, ointments andsprayable liquids or suspensions (U.S. Pat. No. 6,551,577), latexes(U.S. Pat. No. 6,342,212), pastes, oily suspensions, water solublepolymeric films, water-insoluble films capable of sustained release ofthe antimicrobial agent and the like. Additional examples of amorphouscompositions include various types of inks (e.g., flexo, gravure, inkjetinks for DOD and continuous ink jet) and aqueous and non-aqueous resins.

Antimicrobial Active Agents

The antimicrobial compositions of the present disclosure compriseantimicrobial active silver compounds. More specifically, the activecompounds are a group of silver compounds comprising at least ones-triazine ring. Although those ordinarily skilled in the art recognizethat silver containing compounds may have antimicrobial properties, notall compounds are suitable because they may lack the requisite light andheat stability. In that context, antimicrobial silver compoundscomprising s-triazine rings is useful in that they are practically inertto light and heat typically encountered in the handling and/ormanufacturing of antimicrobial compositions and devices. The relevantactive silver compounds comprising the s-triazine ring are described ina published paper (see “Cyanuric acid and cyanurates” by Seifer G. B.,Russian Journal of Coordination Chemistry, Vol. 28, No. 5, p 301-324(2002) which is incorporated in its entirety by reference). Of thosedescribed, example compounds are listed in the table below. Hereafter,the silver compounds listed in Table 1 will be collectively referred toas “silver cyanurate derivatives”.

TABLE 1 Silver cyanurate derivatives of the present disclosure S. No.Silver Compound 1 AgNO₃•C₃N₃ (NH₂)₃ 2 C₃N₃ (NH₂)₂NAg₂ 3 Ammeline•AgNO₃ 4Ammelide•AgNO₃ 5 Mono silver cyanurate (C₃N₃H₂O₃Ag) or its hydrate 6 Disilver cyanurate (C₃N₃HO₃Ag₂) or its hydrate 7 Tri silver cyanurate(C₃N₃O₃Ag₃) or its hydrate 8 Sodium silver cyanurate ligand complexNa[Ag(C₃N₃H₂O₃)₂] or its hydrate 9 Potassium silver cyanurate ligandcomplex K[Ag(C₃N₃H₂O₃)₂] or its hydrate 10 Mixed salt NaAgC₃N₃HO₃ or itshydrate 11 Mixed salt NaAg₂C₃N₃O₃ or its hydrate 12 Mixed saltKAgC₃N₃HO₃ or its hydrate 13 Mixed salt KAg₂C₃N₃O₃ or its hydrate

Of the thirteen example compounds listed, the latter nine (No. 5 to No.13) are highly effective example compounds with (i) Mono silvercyanurate (C₃N₃H₂O₃Ag) or its hydrate, (ii) Di silver cyanurate(C₃N₃HO₃Ag₂) or its hydrate, (iii) Sodium or Potassium silver cyanurateligand complex Na or (K) [Ag(C₃N₃H₂O₃)₂] or its hydrate, (iv) Mixed saltNaAgC₃N₃HO₃ or its hydrate and (v) Mixed salt KAgC₃N₃HO₃ or its hydratebeing particularly attractive for further development.

Although some of the listed compounds have been known for over 175years, those or others have not been suggested for use as antimicrobialagents because those ordinarily skilled in the art recognize that theoverwhelming majority of silver compounds are light and heat sensitiveand therefore reduce easily to elemental silver, which is known to causebrown black discoloration. Moreover, they also recognize there is notheoretical model to predict the degree of light and heat sensitivity ofthe vast number of silver compounds. Furthermore, the light and heatsensitivity of active silver compounds is attenuated in the presence ofvarious other components more so in aqueous environments. Therefore,researching for robust silver compounds is more of an art and yet maybenefit from a more scientific and systematic approach. For example, inthe past forty years, only silver sulfadiazine, silver allantoin complexand to some extent silver saccharinate have been reported as knownantimicrobial silver compounds with some inherent light and heatinsensitivity. But higher insensitivity to light and heat of thesesilver compounds comes at the expense of solubility, which is extremelylow in water. As a result, hydrogel compositions with very high levelsof silver are difficult to formulate without rendering them opaque.

For the example compounds, either of the non-hydrated or hydrated formmay be suitable without adverse effects on antimicrobial activity. Inparticular, one aspect of the present disclosure is that the silvercyanurate derivatives have been observed to be practically inert tolight and heat. Perhaps because these compounds do not reduce easily andthereby resist discoloration by light or heat in an extraordinarymanner. When subjected to intense light or extreme heat, the compoundsare inert as dry solids and are also unaffected as aqueous suspensionsor when dispersed in aqueous amorphous compositions. Thus, an object ofthe present disclosure is to provide antimicrobial silver compounds thatare inert to light and heat as solids or when dispersed in liquids, orin semi-solids such as gels or dispersed in thin solid films such as drycoatings.

Another aspect of the present disclosure is to provide antimicrobialsilver compounds that are sparingly soluble in aqueous or non-aqueousenvironments. These compounds are poorly soluble in water at roomtemperature and therefore release slowly. That allows for prolongedantimicrobial effect in devices and compositions. Due to low solubility,their effective concentration substantially always remains low therebymitigating toxic effects. Yet due to silver's oligodynamic property,these compounds exhibit strong antimicrobial effects but remainnon-toxic to the users. Despite their low water solubility, hydrogelscomprising silver cyanurate derivatives have been formulated up tosilver loadings of 1000 ppm without loss of transparency. For example,the amount of silver in the hydrogel may be between 50 and 1000 ppmbased on a weight of the hydrogel.

Compounds comprising the s-triazine ring such as mono- or dichloroiso-cyanurates are known to have antimicrobial properties and have foundcommercial utility as disinfectants or sanitizers, but they exhibitsignificant toxicity. Yet, cyanuric acid, the starting material for thesilver compounds of the present disclosure is considered relativelynon-toxic and finds use as a stabilizer for N-chloro iso-cyanurates inpool cleaning compositions. According to an internet source (e.g., seewww.wikipedia.org/wiki/Cyanuric_acid) the lethal dose LD50 for rats is7.70 g/kg of body weight. Coupled with low toxicity of silver attherapeutically effective levels, the silver cyanurate derivatives ofthe present disclosure can be considered relatively safe. Furthermore,studies have shown the cyanurates are not metabolized and cleared fromthe human body within 24 h. Thus, another aspect of the disclosure is toprovide antimicrobial silver cyanurate derivatives that are relativelynon-toxic at use levels contemplated for antimicrobial devices andcompositions for topical use.

With regard to the non-toxic nature of cyanuric acid just described(e.g., the lethal dose LD50 for cyanuric acid in rats is 7700 mg/kg ofbody weight), other example acids used in commercial applications arealso known to have higher toxicities. For example, the lethal dose LD50for boric acid in rats is 3450 mg/kg of body weight whereas the lethaldose LD50 for benzoic acid in rats is 2530 mg/kg of body weight; and thelethal dose LD50 for salicylic acid in rats is 1250 mg/kg of bodyweight. Guidelines provided by the US EPA establish toxicityclassifications based on the amount of a substance within a test animal(e.g., rats, fish, mice, cockroaches). An LD50 is a standard measurementof acute toxicity that may be stated in milligrams (mg) of substance perkilogram (kg) of body weight. An LD50 represents the individual doserequired to kill 50 percent of a population of the test animals. Thus,because LD50 values are standard measurements, comparisons may be madeamong various substances using their relative toxicities, and the lowerthe LD50 dose, the more toxic the substance. The LD50 may also be brokeninto additional categories that reflect the type of chemical exposure(e.g., Oral LD50, Inhalation LD50, and Dermal LD50). For example,toxicity classifications (and ranges) for the Dermal LD50 in rats are:high toxicity (LD50≦200 mg/kg); moderate toxicity (200 mg/kg<LD50≦2000mg/kg); low toxicity (2000 mg/kg<LD50≦5000 mg/kg); and very low toxicity(5000 mg/kg<LD50). Thus, with an LD50 of 7700 mg/kg of body weight,cyanuric acid may be classified into the very low toxicity categorybased on the US EPA guidelines provided.

Syntheses of Antimicrobial Silver Cyanurate Derivatives

Another feature of the present disclosure is the relative ease withwhich the silver cyanurate derivatives can be synthesized thereby givingthem an edge over competing products. Typical synthesis conditions aresummarized in the accompanying Table 2. As noted, the silver cyanuratederivatives are obtained simply by combining stock solutions ofingredients in appropriate mole ratios. Due to their poor watersolubility, the compounds precipitate out (typically as white solids) ofthe solutions, are washed multiple times with deionized water to removeside products and unused reactants. If desired the compounds can berecovered as solids after drying for further reformulation work.

TABLE 2 Details of synthesis of various silver cyanurate compounds @20-25 C. Silver nitrate Sodium Order (0.1M) cyanurate of Compound [A](0.1M) [B] addition C₃N₃H₂O₃Ag 1 part 1 part B to A Na[Ag(C₃N₃O₃H₂)₂]0.5 part   1 part A to B C₃N₃O₃HAg₂ 1 part 0.5 part (di sodium) B to ANaAgHC₃N₃O₃ 1 part 1 part A to B C₃N₃O₃Ag₃ 1 part 0.333 part (trisodium) B to A NaAg₂C₃N₃O₃ 1 part 1 part A to BAlternatively, the compounds can be formed in-situ into base amorphouscompositions to introduce antimicrobial functionality. A distinctfeature of their synthesis is the order in which reactants are added; itdictates which of the silver cyanurate derivative form. For instance,when an aliquot of monosodium cyanurate solution is added to an equalvolume of silver nitrate solution of same molarity, monosilver cyanurate(C₃N₃H₂O₃Ag) hydrate is obtained. With the reverse order and the volumeratio of silver nitrate solution to monosodium cyanurate of 0.5, sodiumsilver cyanurate ligand complex Na[Ag(C₃N₃H₂O₃)₂] is formed. Furtherapproaching the ratio of cyanurate to silver ions of 1.0 from a value of2.0, a solid phase rich in silver but with variable composition thatincludes Na[Ag(C₃N₃H₂O₃)₂] is obtained. It is not very clear why suchmixed composition solid phase is formed. But it appears that an excessof cyanurate anions continues to bind to the silver compoundNa[Ag(C₃N₃H₂O₃)₂] already formed giving rise to variable composition.This aspect occurs when mixing monosodium cyanurate and silver nitrate.However, no mixed phase solids result during the syntheses of di-silveror tri-silver cyanurate compounds. While collectively silver cyanuratederivatives are preferred, the solid product with variable compositioncomprising silver derived by adding stock solution of soluble silversalt to the stock solution of monosodium cyanurate in 1:1 mole ratio isalso encompassed by the present disclosure. Similarly, solid productscomprising silver obtained by mixing stock solutions of silver nitrateand ammonium cyanurate in no particular order are also within the scopeof the present disclosure. The molarity of stock solutions of 0.1M inthe syntheses described is for illustration and should not be construedas limiting. In fact the molarities can vary from 0.001 mM to 5.0M.Syntheses of silver cyanurate derivatives have been previously reported(see paper by Seifer G. B. and Tarasova, Z. A., Zh. Neorg. Khim., Vol.34, pp. 1840-43, 1989), which employed 10 mM solutions. The illustrativesynthesis work was performed with monosodium and disodium cyanurate,though similar steps are involved with trisodium cyanurate. Analternative procedure for making trisodium cyanurate was reportedelsewhere (Japanese Patent No. 6829146 as cited in Chemical AbstractVolume 70: P78014 and incorporated here in its entirety by reference).In the syntheses described above, stock solution of silver nitrate wasprepared in deionized water. Thus, the pH was substantially neutral(˜7). However, the stock solution may be kept slightly acidic (pH 2-7range) using an acid, wherein nitric acid is one example. Similarlyslightly alkaline stock solution of alkali metal cyanurates may also beused in the above syntheses. Both of these variations of the preparationof silver cyanurate compounds are encompassed by the present disclosure.

In one aspect of the present disclosure, individual antimicrobial silvercyanurate compounds listed in Table 1 are contemplated for use inpractice. However, they can be either employed singly or as a mixture oftwo or more compounds without departing from the scope of thedisclosure. The use of mixed solid phase compounds obtained when theratio of cyanurate ions to silver ions is between 1 and 2 also fallswithin the scope of the present disclosure. In fact, all silvercyanurate compounds formed as precipitates when the aqueous solutions ofcyanurates (mono-, di- and tri-salt of alkali metals, alkaline earthmetals, barium, magnesium, ammonium, copper, zinc and aluminum) andsoluble silver salt such as silver nitrate are mixed are encompassed bythe present disclosure regardless of whether they may be single chemicalentity or a mixture of various entities. Furthermore, it should berecognized that the use of silver nitrate as a water soluble salt as asource of silver to prepare silver cyanurate derivative is merely forillustrative purposes and is non-limiting. For example, in oneembodiment, silver salts that possess a moderate to high watersolubility at 20-25 C of >5 g/liter is desirable. Though non-limiting,such example silver salts are silver acetate, silver lactate, silvercitrate, silver sulfate and silver phosphate, with silver nitrate beinguseful for development according to the present disclosure.

While the active compounds contemplated for use in the antimicrobialcompositions of the present disclosure are the silver cyanuratederivatives, said compositions may also comprise other silver compounds.These silver salts may or may not be sparingly soluble in water. Forinstance, additional example silver compounds are listed in thepublished US Patent Application No. US2007/0254044 which is incorporatedhere in its entirety by reference. As one example, said compositions mayalso include silver nanoparticles derived by methods disclosed in thepublished US Patent Application No. US2007/0003603. Any one of thesilver cyanurate derivatives in Table 1 and any other silver compoundsfrom those listed in the published US Patent Application No.US2007/0254044 or silver nanoparticles may be used in pairs or asmultiple entity mixtures in hydrogel compositions to provide varyingpatterns of sustained release rates of silver ions for the desiredtherapeutic effect.

In another aspect of the present disclosure is provided the method ofusing alkali metal salts of cyanuric acid. For example, the use ofalkali metals of sodium or potassium though lithium is contemplated.Other metal cyanurates (mono-, di- and tri-) of calcium, magnesium,barium, copper, zinc and aluminum are also encompassed by the presentdisclosure. In addition, the use of ammonium cyanurate and of mixeddi-cyanurates (where the cations are dissimilar) is further contemplatedby the inventor.

Though cyanurates of said metals are known, their commercial applicationhas not been reported. In this context, their use as starting materialsfor a variety of silver cyanurate derivatives becomes a further aspectof the present disclosure. For example, one embodiment of the presentdisclosure is a mixed cyanurate salt of calcium and silver. Such saltcan be used in the manufacture of antimicrobial alginate fibers. Anotherembodiment is the mixed cyanurate salt of barium and silver which can beincorporated into medical devices that provide dual function ofanti-infectivity and of opacity to x-rays.

Mechanistic Aspect of Strong Light and Heat Discoloration Resistance

Without being bound to a theory, the inventor believes the reason behindunprecedented light and heat discoloration observed with theantimicrobial hydrogels comprising silver cyanurate compounds has to dowith the sizes of the crystals of the silver active compounds formedwithin the hydrogels. The crystals are substantially of nano-dimensions(see Table 3 for summary of observations of the crystals of pertinentsilver cyanurate compounds) that may allow the crystals to beintrinsically insensitive to light and heat. Further, the crystals donot readily agglomerate after they are formed perhaps because of thenegative charge on the crystal surfaces. These charges are due to theionization of the hydroxyl groups of cyanuric acid. As all particlescarry like charges, they repel each other which in turn prevents theparticles from coalescing and forming aggregates. The main causes ofdiscoloration is the formation of elemental silver by thephoto-reduction or thermally induced reduction of free silver ions, beso in hydrogel compositions or in simple aqueous solutions. Silver ionsare powerful oxidizers and therefore tend to reduce quickly even inpresence of very weak electron donors. In the present case, withoutbeing bound to theory, the inventor further believes that free silverions concentration in aqueous suspensions of any of the silver cyanuratecompounds in Table 1 is extremely low. This can be deduced from the testobservations (see Example 34) made with aqueous suspensions obtained bymixing monosodium cyanurate and silver nitrate solutions wherein thestarting ratio of silver ions to cyanurate ions was as high as 8. Atsuch a high ratio much in excess of stoichiometry, it was expected thatconsiderable free silver ions would be present. As a result, after steamsterilization, the aqueous suspension would be discolored due to thereduction of free silver ions to Ag⁰. But no such discoloration wasobserved suggesting that the majority of silver was bound to thecyanurate moiety as ligand complex (through N atoms) of unknownstructure. At the same time, because of the retention of antimicrobialactivity, one could deduce that the binding was not strong and was thusreversible.

TABLE 3 Observations on the crystals of silver cyanurate compounds S.No. Compound Water solubility (mg/L) Morphology observed 1 C₃N₃O₃H₂Ag243 Two morphologies seen; majority nanofilaments and nanotrapezoids(<5%); filaments in 3 different sizes, majority 100-500 nm and 1-100 μmlong 2 C₃N₃O₃H₂Ag from Not Tested Single: Nanorods with slant edges;ammonium dominant size: 100-400 nm ans 1-2 cyanurate um long, minoritysize: 400-700 nm and 2-5 μm long 3 Na[Ag(C₃N₃O₃H₂)₂] 1.55 Single;nanofilaments; 100-500 nm dia & 10-100 μm long 4 C₃N₃O₃HAg₂ 851 Single:Nanofilaments or rods; 50-100 nm dia & 500-800 nm long 5 NaAgHC₃N₃O₃1.62 Single: Short nanorods; 200-500 nm dia & 1-2 μm long that appearflat 6 Solid product from Not Tested Two: one amorphous phase withmixing 1 part AgNO₃ pitted surface & nanorods; 1st into 1 part kind300-500 nm dia & 3-5 um Ammonium long; 2nd: 50-200 nm dia & 1-2 μmcyanurate long

Other Actives and Ingredients in Antimicrobial Compositions

The compositions of the present disclosure may also comprise activeagents such as antibiotic and biochemical compounds that may aid and/orpromote wound healing. Non-limiting examples include growth factors,proteins, angiogenic factors, wound healing agents, growth promoters,enzymes, nutrients, vitamins, minerals, mucopolysaccharides, plantderived extracts or chemicals, herbicides, fats, carbohydrates, fattyacids, nucleosides, sera, amino acids, antibodies and fragments thereof,anesthetics, coagulations factors, vesicles with active agents,liposomes with actives including silver, neurochemicals, nitrates,antigens, cellular receptors, metal nanoparticles of silver, gold,copper, zinc, radioactive materials, anti-bacterial agents,anti-microbial agents (chlorohexidine and related compounds, biguanidesand related compounds), anti-viral agents, anti-parasitic agents,anti-fungal agents (azoles and related derivatives), quaternary ammoniumcompounds, indicators of pH, oxidizing agents such as hydrogen peroxide,polyvinylpyrrolidone-peroxide complexes (Peroxydone® K-30 or K-90 ISPCorporation, Wayne, N.J.), organic (urea or melamine or cyanuric acid)and inorganic complexes of hydrogen peroxide (sodium carbonate andvarious other salts) or benzoyl peroxide. Alkali metal cyanurates andsoluble silver salts may be added as dry solids without departing fromthe scope of the disclosure.

In addition to the antimicrobial silver active agent, hydrogelstypically comprise a humectant, a single or multiple viscosity enhancingagent(s) and water as the major component. Optionally, they may comprisea biocompatible coloring agent, skin enhancing additives (e.g.,essential oils, fragrances, moisturizing agents, emollients, toningagents, surfactants etc.) the use of which is known to those ordinarilyinvolved in the topical formulation industry. Optionally, additives suchas buffers to maintain a desired pH are also contemplated for use insaid hydrogels. In this way, the methods described further compriseadding a buffer to the viscous gel and adjusting the pH of theantimicrobial composition to a range of 6 to 8. A variety of options forcoloring hydrogels are also possible. The color may either be impartedby the use of traditional colorant (water soluble dye) or copper-aminoacid complexes. Such complexes are known as the source of absorbent formof copper and find application as dietary food supplements in the animalfeed industry (see U.S. Pat. Nos. 4,900,561 & 7,129,375 which areincorporated here in their entirety by reference). Moreover, FDAapproved dyes for food industry and methylene blue may serve as suitablecolorants. Glycerol, Propylene glycol, polypropylene glycols of varyingmolecular weights and urea are preferred as humectants though otherhumectants such as polyethylene glycols, sodium lactate etc. may also beused. Humectant action may be possible by using one substance or may bederived from two or more. The presence of humectants provides excellentmoisturizing ability to the hydrogel compositions. Therefore, themethods described include making an antimicrobial composition with asilver cyanurate active agent. The method comprises combining aviscosity enhancing agent and a water based solvent to yield a viscousgel, and adding a metal cyanurate solution and a soluble silver saltsolution to the viscous gel, where the metal cyanurate solution and thesoluble silver salt solution react to form the silver cyanurate activeagent. The method further comprises adding a humectant to the viscousgel, where the humectant is one or more of glycerol, propylene glycol,polypropylene glycol, urea, polyethylene glycol, and sodium lactate. Inaddition, the method may further comprise adding a coloring agent to theantimicrobial composition, where the coloring agent is one of a watersoluble dye, a copper-amino acid complex, and methylene blue. In oneexample, the method further includes adding a skin enhancing additive tothe antimicrobial composition, where the skin enhancing additiveincludes one or more of an oil, a fragrance, a moisturizing agent, anemollient, a toning agent, and a surfactant. The gel character of thehydrogels of the present disclosure is brought about by a synthetic claymineral which hydrates when dispersed in water resulting in a very largeincrease in viscosity. One such clay is Laponite® from Southern ClayProducts, Gonzales, Tex. Laponite® clay is the XLG grade. Though, itsuse is for illustration and is non-limiting. Alternately, natural clayminerals or mixtures thereof may also be included in the hydrogels.Other clay minerals that may be used for enhancing viscosity orimparting gel property are disclosed in columns 4 and 5 of U.S. Pat. No.6,333,054 which is incorporated here in its entirety by reference.

To boost viscosity of the hydrogels is not limited to the use of naturalor synthetic clay minerals. Other viscosity enhancing agents orthickeners may be used as well. Non-limiting examples include celluloseethers (sourced from Ashland Chemical Company or Dow Chemical Company orothers) such as hydroxyethyl cellulose, hydroxypropyl cellulose,hydroxyethyl or hydroxypropyl methyl cellulose, sodium carboxymethylcellulose, polyacrylates (sources from Lubrizol Chemical Company andothers), natural gums, chemically modified natural gums, chemicallymodified cellulose ethers with long chain aliphatic chains, syntheticgums, polyvinyl alcohol, polyvinyl pyrrolidone, polyacrylamide,polyaminoacids such as polyaspartate, polyglutamate and2-acrylamido-2-methylpropane sulfonic acid (AMPS) derived polymers. Asone example, polymers may be soluble in water and available in varyingviscosity grades. The polymers may be used singly or as a mixture ofmore than one polymer. In addition, the viscosity increase in saidhydrogels may be derived by using the clay mineral and polymer together.

For the delivery of the active silver, the compositions of the presentdisclosure may comprise hydrogels that have fluid absorbing property.They comprise hydrophilic polymers that swell by trapping water, salineor biological fluids within the polymeric free volume. They may reach anequilibrium absorption capacity (gm of water/gm of dry polymer) thatranges between 1.2 and 300.

The hydrogel compositions contemplated by the present disclosure may beneutral or ionic. A neutral hydrogel is substantially free of anelectric charge and its pH is close to 7. Such neutral hydrogel maycomprise moderate (˜20,000) to high (˜1,000,000) molecular weight PHMMA(polyhydroxyethyl methacrylate), polyvinyl pyrrolidone, polyvinylalcohol with low acetate content, polyethylene oxide polymers withmolecular weights up to 5,000,000, polyvinyl ether polymers; celluloseether polymers with degree of polymerization from 100 to 200,000 such asmethyl cellulose, hydroxyalkylalkyl cellulose derivatives (ethylhydroxyethyl cellulose, hydroxyethyl methylcellulose, hydroxybutylmethylcellulose, hydropropyl methylcellulose, hydroxyl ethylcellulose;neutral polysaccharides such as guar gum, locust bean and tamarind gumand the like. Hydrogels may be made from hydrophilic polymers made frommonomers such as acrylamide, methacrylamide, N-substituted acrylamide,N-substituted methacrylamide and monomethacrylates of polyethyleneglycols.

In contrast, the ionic hydrogel comprises polymers that have chemicalgroups which dissociate in aqueous media and become electricallycharged. The ionic gels may be anionic or cationic. Non-limitingexamples include gels derived from carboxymethyl cellulose polymers,copolymers of maleic acid with styrene, ethylene, propylene, butylene,isobutylene, N-vinyl lactam polymers, polyvinyl sulfonate polymers,phosphorylated hydroxyalkyl methacrylates, Carbopol® brand polymers,polyacrylic acid polymers and copolymers of acrylic acid with acrylamideor methacrylamide, methacrylic acid polymers, anionic derivatives ofcarrageenan, agar, Arabic gum, gum ghatti, and the like. Polymersderived from basic monomers such as aminoalkyl methacrylate, vinylpyridine and other vinyl monomers carrying 5 or 6 member ringscomprising carbon, nitrogen, sulfur and oxygen atoms. In some examples,when preparing hydrogels according to the present disclosure, combiningcationic polymers with Laponite® clay material should be avoided.

A variety of chemical ingredients are suitable as additives in theantimicrobial compositions of the present disclosure. Non-limitingingredients include cellulose ether polymers, sodium alginate, sodiumalginate modified with small amounts of calcium or magnesium ions,propylene glycol or glycerol esters of alginic acid, gum karaya, guargum, gum acacia, gum tragacatha as disclosed in U.S. Pat. No. 4,364,929which is incorporated here in its entirety by reference. Additionalexamples of ingredients include hydratable polyurethane polymers (U.S.Pat. No. 5,175,229), gelatin and its derivatives, naturally occurringpolymers and their derivatives (U.S. Pat. No. 5,804,213), proteinsderived from corn or maize such as zain, hyaluronic acid and derivatives(U.S. Pat. No. 5,128,326), microbial polysaccharides such as beta-1, 3glucan type polysaccharides (U.S. Pat. No. 5,158,772), polyvinyl alcoholderivatives (U.S. Pat. No. 4,708,821), xanthan gum (U.S. Pat. No.4,136,177), locust bean (U.S. Pat. No. 4,136,178) and beta-cyclodextrinderivatives (U.S. Pat. No. 6,468,989), malodextrin and dextrin polymers.

In this way, the method further includes a viscosity enhancing agentselected from one or more of a synthetic clay mineral that includesLaponite®, a natural clay mineral, a cellulose ether selected from thegroup consisting of hydroxyethyl cellulose, hydroxypropyl cellulose,hydroxyethyl cellulose, hydroxypropyl methyl cellulose, and sodiumcarboxymethyl cellulose, polyacrylate, a natural gum, a chemicallymodified natural gum, a chemically modified cellulose ether with analiphatic chains, a synthetic gum, polyvinyl alcohol, polyvinylpyrrolidone, polyacrylamide, and a polyaminoacid that includes one ofpolyaspartate, polyglutamate and AMPS derived polymers. In addition,metal cyanurate solution may include one of sodium cyanurate, potassiumcyanurate, lithium cyanurate, calcium cyanurate, magnesium cyanurate,barium cyanurate, copper cyanurate, zinc cyanurate and aluminumcyanurate. The soluble silver salt solution may further include one ofsilver nitrate, silver acetate, silver lactate, silver citrate, silversulfate and silver phosphate.

With regards to the active agent, according to the methods of thepresent disclosure, the silver cyanurate active agent is one or more ofAgNO₃.C₃N₃(NH₂)₃, C₃N₃(NH₂)₂NAg₂, Ammeline-AgNO₃, Ammelide-AgNO₃,Monosilver cyanurate (C₃N₃H₂O₃Ag), Disilver cyanurate (C₃N₃HO₃Ag₂),Trisilver cyanurate (C₃N₃O₃Ag₃), a sodium silver cyanurate ligandcomplex Na[Ag(C₃N₃H₂O₃)₂], a potassium silver cyanurate ligand complexK[Ag(C₃N₃H₂O₃)₂], a mixed salt of NaAgC₃N₃HO₃, a mixed salt ofNaAg₂C₃N₃O₃, a mixed salt of KAgC₃N₃HO₃, a mixed salt KAg₂C₃N₃O₃, and ahydrated species thereof.

The water based compositions of the present disclosure particularlythose suitable for medical purposes may comprise viscosities from about1.0 centipoise (cP) to about 2,000,000 cP at 25 C and preferably fromabout 1 to 1,000,000 cP as measured on the Brookfield Viscometer. Forthose compositions that exhibit thixotropy, the yield stress asdetermined from frequency and strain sweep data may be about 1.0 Pascal(Pa) to about 10,000 Pa at 25 C.

The antimicrobial silver compositions, both aqueous and non-aqueoustargeted for industrial uses may actually possess viscosities from about0.0001 cP to about 2,000,000,000 cP at 25 C.

The water employed to produce hydrogels may be either deionized water ordistilled water. Though desired that the water be pyrogen free,especially for medical purposes, it is not required. The non-aqueouscompositions comprise non-aqueous solvents as a major constituent.Suitable examples include, acetone, methyl ethyl ketone, lower alkylalcohols (C₁-C₆ carbon atoms) and their esters, cellosolve typesolvents, THF, DMSO, DMF, propylene glycol, ethylene glycol, toluene andC₅-C₁₀ alkanes.

While the hydrogel compositions may be thixotropic, they also may or maynot be thixotropic or clear or colorless or opaque. All such variationsare encompassed by the present disclosure. While thixotropic characteris a qualitative measure, the yield stress derived from establishedrheological measurement is quantitative and therefore represents a morerobust metric of performance. In this way, hydrogels of the presentdisclosure possess yield stress values between 0 and 1000 Pa at 20-25 C,further between 5 and 750 Pa and further between 50 and 400 Pa. Besideshydrogels, the antimicrobial compositions of the present disclosure maytake a variety of forms. They may be o/w emulsions or w/o emulsions or“emugels”, salves, pessaries, jellies, creams, lotions, plain solutionsand suspensions and may be so formulated to meet different customerrequirements.

One embodiment of the present disclosure is an antimicrobial petroleumjelly with a silver content of ˜540 ppm. It was derived by blending intoplain petroleum jelly (Vaseline® brand obtained from a local store) asmall aliquot of the suspension of silver cyanurate compound. Thesuspension of silver cyanurate compound was prepared by adding silvernitrate to monosodium cyanurate solution in 1:1 mole ratio, spiked witha small quantity of an emulsifier (e.g., Tween® 20, Sigma Aldrich). Theresulting antimicrobial composition was smooth, opaque white, withstoodcontinuous light exposure for a week without signs of discoloration andexhibited a zone of inhibition against Staphylococcus aureus ATCC 6538in a ZOI assay. Such a composition could provide a sustained sanitizingeffect compared to the alcohol based hand sanitizers and without theexcessive alcohol induced dry feel.

Another example of antimicrobial composition of the present disclosureis a simple suspension of silver cyanurate compound. The suspension wasprepared by simply mixing silver nitrate and monosodium cyanuratesolutions (0.1M) in 1:1 volume ratio. When the molarity of the solutionsis kept low, the resulting suspensions are actually clear and gives ahint of pearlescence. Moreover, these suspensions can be steamsterilized, are inert to light induced discoloration, have pH nearneutral and have a practically indefinite shelf life given they areimmune to environmental temperature changes. The ability to withstandsteam sterilization cycles without a loss of antimicrobial effectivenessin aqueous medium and retention of its native white color distinguishthe inventive compounds from the rest of the silver active compounds.This property is another feature of the present disclosure.

In an embodiment of the present disclosure, the suspension or solutionsmay be used as cleaning or sanitizing solutions in hospitals, ERs,surgical suits and in wound care. In yet another embodiment, theantimicrobial compositions (sterilized solutions or suspensions)comprising silver cyanurate compounds could be paired with a sterileabsorbent gel sheet similar to a Flexigel® sheet as a wound care kit.The antimicrobial composition can be packaged in dark glass ampoules (asa precaution) and then steam sterilized. The gel sheet can be pouched infoil packaging and sterilized by e-beam or gamma irradiation. When usedto cleanse the wound to reduce the bio-burden, the glass ampoule isbroken and the sterile antimicrobial composition is poured into thepouch, left for few minutes to soak up the fluids. The now hydrated gelsheet is then removed from the pouch and applied to the wound that isbandaged up. In a modification of the embodiment disclosed here, the gelsheet could be replaced by an absorbent polyurethane sheet. Such foamsare widely available commercially, e.g., from Rynel Inc. and LendellManufacturing Company, a unit of Filtrona Corporation, St. Charles,Mich. Polyurethane foams comprising various silver active agents(nanosilver, silver saccharinate) have been disclosed in the patentliterature (US 2012/0322903 which is incorporated here in their entiretyby reference). A Polyurethane foam comprising silver cyanurate compoundcan be prepared by following the procedure described in the US publishedPatent Application No. 2012/0322903 with small modifications. Forinstance, one can omit the solubilization step because the silvercyanurate compounds in suspension form as extremely fine particulatesbordering on a colloidal state. Sterile solutions for treating earinfections represent another application where the simple liquidcompositions of the present disclosure may be useful. The sterilesolutions of silver cyanurate derivatives may also be used forsterilizing all types of orthopedic devices prior to implantation andmay be an adequate substitute for the antibiotic cocktails currentlyemployed. Furthermore, the sterile solutions or liquids comprisingsilver cyanurate derivatives may also be used to disinfect dental toolsand may be suitable for use in dental implants to provide antimicrobialenvironment and prevent the growth of foul odor causing anaerobes inhard to reach gaps.

Another area where the liquid compositions of silver cyanurate compoundsmay find use is in negative pressure wound therapy (NPWT). Prior to usein NPWT, the sterile foam block may be wetted with sterile liquidscomprising silver cyanurate derivatives. When in contact with the wound,silver in the liquids may rapidly diffuse to its surroundings, therebykilling the bacteria and reducing bio-burden. In this way, it may helplower the bio-burden, but it may further help control foul wound odors.

In yet another application of the liquid compositions comprising silvercyanurate compounds, they can be used in contact lens maintenance. Anaqueous concentrate of the active compound or mixture thereof may besterilized in a polypropylene bottle. A few drops of the concentrate maybe added to the lens maintenance solution in the lens case to killbacteria and prevent their growth. Due to silver's broad spectrumantimicrobial property it is also effective against amoeba some of whichhave been implicated in outbreak of infections among contact lens users.In another alternate application, the sterile solutions of the silvercyanurate derivatives may be used to disinfect drinking water. Thisapplication is more suited to emergencies or in developing countries. Afew drops of aqueous concentrate comprising silver may be added to waterto disinfect it and make is potable. Due to very low toxicity ofcyanurate moiety, the risk to the users may be minimal and the benefitof using the same may far outweigh the potential harm caused byingesting dirty water. The compounds can be added to liquid mixturescomprising water or non-aqueous solvents, o/w or w/o or w/o/w emulsions,emulgels, gels or suspensions. The fluid compositions may be used fordisinfection of walls, floors, counters and table tops. It may beincorporated into cosmetics and surgical scrubs.

Another embodiment of antimicrobial aqueous compositions are water basedpaints such as acrylic paints. These paints include acrylic polymersthat are normally water insoluble. Under alkaline pH, the acrylic acidgroups of the polymers are converted to their ammonium salt. Into thesepaints, the silver cyanurate compounds can be formed in-situ or added asdry fine solids to impart antimicrobial function. Interestingly, not allof these active compounds are affected by pH >7. When surfaces arepainted with such paints, the ammonia is lost to the atmosphere thusreturning the acrylic polymer to insoluble antimicrobial paint layer.Such paints could be useful to keep the walls in operating rooms inhospitals germ free. Oil based paints may also incorporate the silvercyanurate derivatives without departing from the scope of the presentdisclosure.

A modification of the embodiment of antimicrobial paint is anantimicrobial sodium silicate composition comprising silver cyanuratecompounds. In preparing such a composition, a suspension of silvercyanurate compound was blended into 40% by weight aqueous sodiumsilicate solution to obtain a milky viscous mixture. To simulate a reallife application to a tiled surface, a glass slide was coated with themixture that was cured at 110 C to form insoluble glass with finelydivided trapped particles of the active compounds. The coating did notdiscolor after 30 days of continuous light exposure and exhibitedantimicrobial activity in a ZOI assay.

In another embodiment, the silver cyanurate derivatives may be includedin creams to treat diaper rash. As an example, a commercial diaper rashointment was blended with silver cyanurate compound in a manner similarto that described earlier to prepare petroleum jelly. There was novisual difference between the ointment samples with and without silver.Though in ZOI assay, the ointment with silver showed a large inhibitionzone consistent with its increased antimicrobial potency.

Another aspect of the present disclosure is to provide inert solidsubstrates of varying contents of silver cyanurate compounds. Suchsubstrates may be powders both natural and synthetic or made ofinorganic porous supports, ceramics, metals, oxides, pellets, shortfibers etc. In preparing one such embodiment, talc powder was dispersedin a mixture of water and ethanol. To the suspension formed, monosodiumcyanurate solution was added followed quickly by equal aliquot of silvernitrate solution to evenly precipitate out an insoluble fine silvercyanurate derivative compound. The talc powder with dispersed activecompound was recovered after filtration, washing and dried. Visually, nodifference was observed between dry talc with and without silver. Suchsilver impregnated talc could be convenient to use in the treatment ofathlete's foot or to fight foot odors caused by bacteria. They may bealso suitable for use by diabetics to prevent wound infection on feet.

Antimicrobial Medical and Non-Medical Devices

A large number of devices for use in medical industry and non-medicalindustry can be rendered antimicrobial using the amorphous compositionscomprising the silver cyanurate derivatives. For example, coatings maybe applied to surfaces of pre-made three dimensional objects or articlesby traditional means such as dip coating, brushing or spraying. As such,the antimicrobial compound may be blended into a surface coating of amedical device. In other cases, the actives may be incorporated intopre-mixes and then the objects or articles molded into shapes. Suitabledevices that can be imparted antimicrobial function by the silvercyanurate compounds of the present disclosure include air and waterfiltration devices, air ductwork, fan housings, aquarium filtermaterial, automobile ventilation, air conditioner systems, bed sheets,blankets and bed spreads, buffer pads (abrasive and polishing), carpetsand draperies, fiberfill for upholstery, sleeping bags, apparel, etc.where the fiber is cellulose (natural or regenerated), natural down,nylon, polyester, rayon, wool, fiberglass duct board, fiber hose fabric,humidifier belts, mattress pads, underwear and outwear, nonwovendisposable baby and adult diapers, tampons, nonwoven polyester, campgear apparel, PU foam cushions, PU foam for household, industrial andinstitutional sponges and mops, PU foam for packaging and cushioning, PUfoam as growth medium for crops and plants, pre-moistened towelettes andtissue wipes, roofing materials such as shingles, roofing granules, woodshakes, wood planks of various widths, lengths and sizes, felt, stoneand synthetic overcoats, sand bags, tents, tarps, sails and ropes,athletic, casual and dress shoes, shoe insoles and inserts, leather andleather like products, shower curtains, socks for athlete's foot funguscontrol, throw rugs, towels made of 100% cotton or polyesters or theirblends, toilet tank, toilet cleaning tablet and seat covers, umbrellas,upholstery made of acetates, acrylics, cotton, fiberglass, nylon,polyester, PE, polyolefins, PP, rayon, spandex, vinyl and wool, vacuumcleaner bags and filters, vinyl or wall papers, disposable wiping clothsfor dusting and washing furniture, car, walls, windows, doors,appliances, dishes, counter tops etc. women's hosiery and women'sintimate apparel. Additional industrial items include food packaging,drug and cosmetic packaging, eating utensils, shower curtains, bath matsand the like, compositions such as grout, cement and concrete to fightgrowth of mold and mildew, sponges, toilet seats, kitchen, bath or labshelf paper, carpet pads, preservative packets for flower bouquets thatmay as one example be used to prevent and/or limit microbial growthwithin the bouquet, pool covers, solar pool covers, cat litter, animalbedding, individual computer keyboards and replacement keys, door knobs,tampons, sanitary napkins, dental chairs, dry sheets, mops anddishcloths, adhesives, silicone products (tubing, plugs, sheets etc.),and microbeads made from natural and synthetic polymers and friablebeads. In this way, the antimicrobial compounds may be useful innon-sterile as well as sterile applications.

The medical devices suitable for imparting antimicrobial propertyutilizing the compositions comprising silver cyanurate derivativesinclude catheters, blood lines, metal or metal alloy implants andorthopedic devices, prosthetic devices and inserts, thermometers,bandages, surgical dressings, surgical apparel, face masks, respirators,wound care and ostomy products, rubber gloves, contact lenses, hearingaids, implantable hearing devices and dusting powder. Examples of fiberand fabric products contemplated include, but are not limited tosurgical gauze, padding on wound dressings, mattress covers, cribcovers, bassinet covers, sailboat sails, tents, draw sheets, cubiclecurtains, tooth brushes, hair brushes, fabric wall coverings, fabricbase, fabric shower curtains, bath mats, athletic clothing such asunderclothes, shirts, socks, shorts, pants, shoes, hospital clothingsuch as examination robes, physicians coats and nurses uniforms, bloodpressure measuring device etc. Additional examples of both medical andnon-medical devices that can be rendered antimicrobial with the silvercyanurate derivatives of the present disclosure are listed in para [140]of US published Patent Application No. US 2007/0003603 which isincorporated here in its entirety by reference.

The silver cyanurate derivatives may be applied topically to both,natural or synthetic fiber or incorporated directly into the syntheticfibers during fiber manufacturing process. The fibers are not limited towool, cotton, polyolefins, polyester, polyamides, cellulose acetate,rayon, polystyrene, vinyls, acrylics and PU's.

The antimicrobial medical and non-medical devices comprising the silvercyanurate compounds of the present disclosure may be sterilized by theknown methods such as ETO, steam sterilization, E-beam and gammairradiation. Even the antimicrobial amorphous compositions besides steamsterilization may be sterilized by E-beam and gamma irradiation at verylow KGy dosages.

Methods of Making the Antimicrobial Compositions

Another aspect of the present disclosure include methods of making theantimicrobial compositions comprising the silver cyanurate compounds.One method comprises the steps of combining a viscosity enhancing agentor thickener, a humectant with deionized water to yield a viscous geland adding successively to said gel metal cyanurate solution and asoluble silver salt solution to the active silver cyanurate compoundin-situ. The step involving the soluble silver salt solution addition iscarried out in the dark though it may be carried out under low lightingconditions without departing from the scope of the disclosure. Theresulting composition is an antimicrobial hydrogel with pH inphysiological range (6-8) that is thixotropic, spreadable, smooth, clearor transparent and moisturizing. Furthermore, the hydrogel and itsvariants have low to moderate capacity to absorb additional water whichassists in the moisture management of the wounds. In another method toprepare said hydrogel, active silver cyanurate compound is pre-made as amilky white suspension by mixing appropriate volumes of equimolarmonosodium cyanurate and soluble silver salt solutions and then blendedin.

Some ingredients such as thickeners and humectants used in thepreparation of said compositions are commercially available. Of thosethat are cosmetic grade or FCC/NF/USP grade are attractive for use. Asto the source of the active silver, ACS grade soluble silver salts maybe used though the high purity USP grade may also be used. Variousalkali metal cyanurate compounds which are not commercially availableare synthesized using cyanuric acid. First, commercial grade cyanuricacid is purified to remove acid soluble impurities followed byneutralization reaction with sodium hydroxide. When the acid and baseare in 1:1 mole ratio, monosodium cyanurate is obtained which isrecrystallized from water for further purity. Presence of additionalsodium hydroxide yields di-sodium cyanurate. One can obtain tri-sodiumcyanurate using large excess of sodium hydroxide as reported in thepublished literature. Similarly, one can obtain ammonium cyanuratecrystals by simply combining a slight molar excess of ammonium hydroxidewith cyanuric acid. In the preparation of alkali metal and ammoniumcyanurate salts, it is desirable to heat the reaction mixtures to 80-85C for 0.25 to 4 hours, however heating for 0.25-2 hours may also be usedto drive the reactions to completion.

Various embodiments of the methods described above are possible witheach of them encompassed by the present disclosure. For instance, in oneembodiment of the method, the humectant is added in the last step. In asecond embodiment, the thickener or viscosity enhancing agent and thehumectant may be combined to from a viscous blend that is then hydratedwith water and then the active compound is formed in-situ in the finalsteps. In a third embodiment, the active agent suspension is preparedand then diluted with water. The thickener e.g. Laponite XLG is hydratedinto the diluted suspension and finally the humectant is added. Thougheach of the silver cyanurate compounds of the present disclosure areantimicrobial, which of them are formed in-situ in the hydrogelcompositions or in suspensions is simply dictated by the order ofaddition of the reagent solutions and their volume ratio. In general,the hydrogel preparations of the present disclosure may be implementedwith a wide range of the molarity of the metal cyanurates or solublesilver salt solutions. Thus, the molarity of the said solutions can bebetween 0.001 mM and 5M though values between 0.001M and 0.5M are alsopossible, and values between 0.05M and 0.2M are further possible.

In general, the methods of making hydrogels of the present disclosurecontemplate the formation in the composition of one kind of activesilver cyanurate compound. However, a method to make hydrogel with twoor more active silver cyanurate compounds is not outside the scope ofthe present disclosure. For example, to the base gel comprising thethickener and humectant in water, one may add a suspension ofmono-silver cyanurate and a suspension of di-silver cyanurate therebyderiving a hydrogel with two silver cyanurate actives. Optionally, othersilver salts solutions may be added to yield different hydrogelcompositions. Alternately, two or more actives may be formed in-situ inthe base hydrogel (composed of thickener, humectant and deionizedwater). To those ordinarily skilled in the formulation industry, it willbe apparent that a large number of permutations and combinations of theactives are possible with each considered within the scope of thepresent disclosure.

In another inventive modification of the hydrogel preparation, the useof metal cyanurate is omitted. In obtaining the finished hydrogel, themethod comprises steps of (i) dissolving cyanuric acid in deionizedwater, (ii) dispersing and hydrating Laponite XLG clay to the acidicwater, (iii) adding soluble silver salt solution in an amountcorresponding to 1:1 mole ratio with the said acid and finally, (iv)adding the humectant.

In another embodiment of the method of making hydrogel composition, twothickening agents are separately dissolved in deionized water and thenthe solutions combined, followed by the humectant, and precipitationin-situ of the active silver compounds. The advantage of this embodimentis the reduction in the opacity of the finished hydrogel especially ifone the thickening agent is a synthetic clay Laponite XLG.

The employment of the silver cyanurate compounds is not limited toimparting antimicrobial property to just hydrogel compositions. Thesecompounds can efficiently be incorporated into aqueous and non-aqueouscompositions, devices, objects and substrates such as paper and fibers.

One embodiment of the aqueous composition comprises a single activesilver cyanurate compound suspended in water. Optionally the aqueouscomposition may include a biocompatible polymer such as polyethyleneoxide polymer or polyvinyl alcohol of low to moderate molecular weight(MW: 20,000 to 200,000) to stabilize the suspension and prevent denseparticles of the active silver compound from settling. The use ofsurfactants such as the Tween® or Span® family surfactant in thecompositions to increase compatibility with hydrophobic constituents isalso contemplated by the inventor. The amount of silver present in suchcompositions may be between 0.0001% and 1.0% by weight, but may also bebetween 0.002% and 0.8% by weight and may further be between 0.0025% and0.5% by weight. The amount of polymer or surfactant can be between0.001% and 10% by weight and but may also be between 0.005% and 1.0% byweight. The preferred polymer grade is USP. Other polymers such ascellulose ether polymers or polyvinyl pyrrolidone may also be used insuch compositions and their use is within the scope of the presentdisclosure.

The following describes a method of making an embodiment in the form ofa w/o emulsion or cream, where the oil phase is petroleum jelly andwater phase comprises active silver cyanurate compound. The methodcomprises the steps of (i) preparing a suspension of the active silvercyanurate compound by admixing solutions of soluble silver salt andmonosodium cyanurate, (ii) further adding to the suspension anemulsifier Tween 20 and dispersing the aqueous mixture into thepetroleum jelly to produce whitish opaque cream. The same method may beemployed to prepare a cream that may also comprise zinc oxide.

In an aspect of the present disclosure, a method is provided to impartantimicrobial properties to a paper substrate. The method may also beapplied to render woven or non-woven fibrous material (derived eitherfrom natural or synthetic sources) antimicrobial especially those thatare wetted by water or mixtures of non-aqueous solvents such as acetone,THF, and alcohols with water. The method comprises the steps of (i)adding soluble silver salt solution to mono sodium cyanurate wherein thecyanurate anion is in excess of silver ions, (ii) diluting the resultingsuspension of the active silver cyanurate compound with dilute aqueousammonia, (iii) dipping the non-woven substrate for time sufficient toallow for the substrate to absorb the fluids, (iv) squeezing out excessfluid and finally (v) drying the substrate to remove all residualsolvents. In a variation of the above method, in the step (ii) insteadof dilute ammonia one may use a non-aqueous solvent or a mixture ofwater miscible non-aqueous solvent and water wherein the non-aqueoussolvent is more than 50% by volume.

The methods of making antimicrobial compositions of the presentdisclosure are quite versatile to implement. By simply adjusting themolarity and or the volume of soluble silver salt solution utilized onecan tailor the desired amount of silver loading in the composition.Moreover, by selecting the order of addition of the two reagents—silversalt and alkaline cyanurate solution—one can choose the type of activesilver compound desired in the composition. Further, one may incorporatea variety of active silver compounds, all with excellent antimicrobialeffects simply by choosing either a mono-, di- or tri-sodium cyanurateor any other metal cyanurate as the anion exchanging compound.

Methods of Using the Antimicrobial Compositions

In one embodiment of the method, the hydrogel may be used to treattopical infection. The composition is generously applied to the infectedarea of the skin and appropriately to the surroundings and then coveredwith a dressing. In a modification of the method, the hydrogel withmoderate level of silver may be used to treat infection in partialthickness or deep wound. The hydrogel in a quantity sufficient to betherapeutically effective is applied to the wound and the surroundingarea and covered with a dressing. Optionally a sterile absorbent foamsheet dressing may be applied over the hydrogel to enhance exudateabsorption. In another embodiment of the method, the hydrogels may beused to treat pressure ulcers, partial and full thickness wounds,diabetic foot and leg ulcers, graft and donor sites, and first andsecond degree burns. In yet another embodiment, the hydrogels with highlevels of silver content may be used to reduce the bio-burden ofgangrenous wounds followed by the use of low silver hydrogels tomaintain the wound bacteria free and accelerate the wound healingprocesses.

A related embodiment of the method of treating infected wounds comprisesa kit that includes a sterile antimicrobial aqueous suspensioncomprising silver cyanurate compounds (contained in an ampoule) of thepresent disclosure and a sterile hydrogel sheet contained in a pouchcapable of absorbing fluids when contacted with infected wounds and themethod of using the kit to treat infected wounds. The treatmentcomprises the steps of (i) breaking the ampoule and opening the pouch byaseptic means, (ii) combining the antimicrobial aqueous suspension bypouring into the pouch, (iii) maintaining intimate contact between thesuspension and the said hydrogel sheet for sufficient time to absorb thefluids partially and coat the antimicrobial active on the said hydrogelsheet, (iv) removing the partially hydrated hydrogel sheet and placingit over the infected wound and finally (v) applying a dressing to coverthe wound. Examples of hydrogel sheet suitable for use are Flexigel® andGeliperm® brand sheets. The method embodied in the invention may alsoutilize sterile non-woven alginate dressings or a hydrophilic PU foam orcotton gauze that do not possess antimicrobial property withoutdeparting from the scope of the present disclosure. The duration betweendressings changes in practice and may be dictated by how long theantimicrobial effect is sustained, which in turn depends on the silverloading of the aqueous suspension. In one embodiment, silver loadingsare those that sustain antimicrobial effect for three to seven days.

In hydrogel compositions wherein the silver loading levels are low, theantimicrobial effect may last a day or two. Such hydrogel compositionsmay be used to treat minor skin cuts or abrasions or very small areaburns. For instance, a hydrogel bead is applied to the cut or burn andthen covered with a bandage strip. Based on the ability of the hydrogelcompositions of the present disclosure to resist light and heat induceddiscoloration, they may be applied on the skin without the risk ofstaining.

In yet another embodiment of the method, said hydrogel compositions maybe used to treat dermal conditions such as acne, rosacea, jock itch,athlete's foot and onychomycosis (nail fungus infection) which arecaused by a host of micro-organisms. The broad spectrum silvercyanurates are effective against causative agents for these dermalconditions, namely the anaerobic bacteria (acne), demodex mites(rosacea), fungi (jock itch and athlete's foot) and dermatophytes(onychomycosis). The silver content of antimicrobial compositions fortreating acne, rosacea, jock itch, athlete's foot and onychomycosis ispreferably between 0.01% and 0.3% by weight. To be effective in treatingthese dermal conditions, said compositions are applied evenly to acnepimples or the affected area in the case of rosacea and jock itch orathlete's foot. The affected nail bed is evenly covered with saidcompositions spread as a layer and covered with a dressing. Thetreatment durations in practice may vary depending upon the severity ofthe respective condition.

In another embodiment of the present disclosure is provided a method ofpreventing or inhibiting biofilm formation on a surface. The methodcomprises the steps of (i) preparing a coating solution comprising oneor more active silver cyanurate compound, (ii) applying said coatingsolution to the surface, and finally (iii) drying the coating to removesolvent residues. The coating solution can be water based or can be madeof non-aqueous solvents or mixtures and prepared by dissolving suitablepolymers and adding a suspension of the active silver compound orforming the active compound in-situ. Any suitable hydrophilic polymermay be employed, including, for example, polyhydroxyethyl methacrylate,polyacrylamide, polydimethylsiloxane, N-vinyl-2-pyrrolidinone,hydrophilic polyurethane, and the like. The hydrophilic polymer may behydrophilic polyurethane, such as the TECOPHILIC™. polyurethane sold byThermedics of Woburn, Mass., for example. Examples of lipophilicpolymers include silicone, polyurethane, polyethylene, nylon, polyvinylchloride, polyvinyl alcohol, the cellulosic polymers, polyvinyl acetate,polyesters, and acrylics. For implants, the coatings are derived frombierodable polymers or bioabsorbable polymers which are known to thoseskilled in the art.

In another aspect of the present disclosure, the hydrogel compositionscomprising silver cyanurate compounds with high silver content may serveas anti-viral compositions to treat and heal cold sores caused by herpessimplex virus. The compositions may be applied in small amounts to covereach sore. The active silver may neutralize the virus by attaching toviral proteins and thereby reduce its infectivity. In addition, it mayalso aid in healing the sore quickly by disinfecting the area. Theantimicrobial hydrogel compositions for antiviral application comprisesilver in the range of 0.005% and 5.00% by weight, however silver in therange 0.01% and 3.00% by weight may also be used, and silver in therange 0.1% and 0.5% by weight may be further used in some examples.

In another embodiment of the methods of the present disclosure, thehydrogel compositions are used to reduce the risk of infection to womendue to HIV during sexual intercourse. Though, it has been reported thatsilver nanoparticles in concentration of ˜1000 ppm can inhibit HIV,ionic silver present as salts was not found to be as effective. Theeffectiveness of silver nanoparticles against HIV inactivation wasattributed to the nano dimensions of the particles that allowed theirgreater interaction with HIV. In contrast, because it is difficult tomaintain ionic silver at high concentration in suitable vehicles such asgels as it deactivates rapidly by photo-reduction or is reduced by heat(visual indication is that it turns black or grey) previous silvercompositions were ineffective. The antimicrobial hydrogel compositionscure this deficiency as they possess excellent stability againstdeactivation. In addition, the active silver cyanurate compounds in thehydrogel compositions have been observed by high resolution SEM topossess nano-dimensions that increase the probability of their lethalinteraction with viruses. Furthermore, the hydrogel compositions aresmooth, viscous, thixotropic, transparent, have pH in physiologicalrange and are readily spreadable under shear forces generated in topicaluse and are convenient to use as a vaginal lubricant. As such, thehydrogel compositions may be applied to an uninfected individual toreduce the risk of infection due to HIV during sexual contact.

In one related embodiment of the disclosure, said compositions areprovided in convenient single use disposal packets and can withstand theharsh environmental conditions of third world countries located in Asiaand Africa. To reduce the risk of HIV transmitting to an uninfectedfemale during sexual contact, said compositions are applied insufficient amount to cover the vaginal area including folds prior tosexual contact. The risk of infection during sexual intercourse may bereduced as the silver may inactivate viral particles by attaching toelectron donating groups present on viral proteins and so prevent viralreplication.

In yet another embodiment, the silver cyanurate compounds of the presentdisclosure may be very fine crystalline materials having nano-sizedimensions. Thus, their ease of preparation and uniformity andconsistency of crystal morphology may further enable their use insecurity applications as taggants. For example, the compounds can beradioactive and so derived from the combination of a metal cyanuratewith a radioactive silver nitrate (^(110m)AgNO₃). As these compounds arealso inert to heat and light, they may be quite robust as security tagsor shelf life indicators for time sensitive products (e.g., productswith a shelf life of 6 to 9 months shelf life).

Ranges of Ingredients in the Antimicrobial Compositions and Devices

The amount of silver in the hydrogel compositions of the presentdisclosure may vary between 0.005% weight and 5.00% weight. However, insome embodiments, a range of 0.005%-2.5% weight may also be used.Further, in other embodiments, a range of 0.01%-0.50% weight may also beused. In this way, the amount of silver in the non-hydrogelantimicrobial compositions and devices ranges between 0.0001% weight and5.00% weight. In still other embodiments, a range may be determined inparts per million (or ppm) based on a composition. Thus, in anon-hydrogel, the amount of silver in a silver cyanurate active agentmay be between 10 and 5500 ppm based on a weight of the antimicrobialcomposition. Moreover, a different range may be used based on the typeof application. Thus, in a hydrogel, the amount of silver in thehydrogel may be between 50 and 1000 ppm based on a weight of thehydrogel.

The amount of thickener in the said antimicrobial hydrogel compositionsmay vary between 0.10% weight and 10.00% weight, though the range0.25%-7.50% weight may also be used. If two thickeners are employed, theweight ratio of one thickener to the second thickener may vary fromabout 1:20 to about 20:1 with total thickener content restricted by theabove range. The humectant concentration of said antimicrobial hydrogelcompositions may vary between 1.00% weight and 40.00% weight though therange 5.00%-20.00% weight may also be used. If two humectants are used,their weight ratio similarly may vary from about 1:20 to 20:1. The totaladditives content (including colorant, skin enhancing agents etc.) ofsaid antimicrobial hydrogel compositions may vary between 0.0001% weightand 5.00% weight. The minimum amount of water in said antimicrobialhydrogel compositions is 40.00% weight and is adjusted once theconcentrations of other ingredients are fixed. This minimum amount ofwater does not apply to other non-hydrogel antimicrobial compositionssuch as suspensions, o/w emulsions, w/o emulsions or emulgels, pastes,oily suspensions or liquids or other non-aqueous amorphous compositions.The amount of silver in the antimicrobial devices of the presentdisclosure may vary between 0.0001% weight and 10.00% weight based onthe weight of the device.

Test Methods

Various test methods were attempted to evaluate the robustness of theantimicrobial compositions and devices of the present disclosure.

Light Exposure Testing

(a) Table lamp light exposure test (TLE)

-   -   The samples of the said compositions contained in either glass        vials or 15 ml PP tubes (BD Falcon) were placed under a table        lamp (turned on) at a distance of 12 to 15 inches for continuous        exposure. The incandescent lamp wattage was 60 W. After the        desired duration of exposure, the test samples were examined for        visible discoloration by holding it against a white plain paper.        Non-hydrogel samples were examined against the control samples        protected from light.

(b) Sun light exposure test (SLE)

-   -   The samples of the test compositions were contained in either        glass vials or 15 ml PP tubes and exposed to direct sunlight.        The exposure was carried out during the hours of 9:00 am and        3:00 pm., and the exposure testing took place over the calendar        year. The intensity of sunlight corresponded to the sunlight        experienced at 45N latitude.

Thermal Testing

(a) Accelerated age test

-   -   To assess the prototype hydrogel compositions for shelf life,        samples were placed in 15 ml PP tubes or were contained in        commercial PE or PP tubing and placed in an oven set to 55 C.        The samples were visually examined qualitatively for        discoloration or physical changes such as loss of viscosity and        syneresis.

(b) Steam sterilization

-   -   One steam sterilization cycle was imposed on gel prototypes to        evaluate their ability to withstand elevated temperatures. The        gel samples contained in 15 ml or 50 ml PP tubes or other        non-hydrogel prototypes (in sealed foil pouches) were examined        post sterilization for any adverse temperature effect on its        color, viscosity, texture, phase separation etc.

Microbiological Testing

The antimicrobial activity of the hydrogel compositions and variousdevice prototypes comprising silver cyanurate compounds was verified bystandard zone of inhibition assay known to those skilled in the art.Briefly, in this assay, samples were placed on plates with proprietaryagar formula (similar to Mueller Hinton Agar (MHA)) that were inoculatedwith bacteria and incubated at 37 C overnight. If antimicrobial activityin the sample was present, it formed a clear zone around the edges. Asnegative control, the samples without the silver active compound wereused. In some tests, positive control was provided by use of commercialproduct samples with silver active compounds, e.g. Silvasorb®, Normlgel®Ag, SilverSept® gel or Maxorb® Ag. Primarily two microorganisms, one agram positive bacteria Staphylococcus aureus ATCC6538 and the other gramnegative Pseudomonas aeruginosa ATCC9027 were employed in the assay. Toexamine broad spectrum antimicrobial activity, various different typesof organisms including MRSA and VRE were obtained. In investigating thebroad spectrum activity, the ZOI assay was performed slightlydifferently. Instead of laying the samples on plates individuallyinoculated with different types of bacteria, the bacterial inoculumswere streaked as parallel lines on one plate. After streaking theinoculum linearly, the samples were deposited as a continuous beadstring in perpendicular direction to the streak lines. Evidence ofantimicrobial activity in the sample was seen in the form ofinterruptions on both sides of the edges of the sample string.

A bacterial challenge assay was employed to verify antimicrobialactivity of liquid compositions (suspensions) of the silver cyanuratecompounds. Briefly, to the aliquots of the liquid compositions,bacterial inoculums were added and the samples were incubated at 37 Covernight. Thereafter, the test samples with silver were treated withsodium thioglycolate solution to neutralize silver and plated on agarplates (note if the samples contained less than 75 ppm silver, the useof sodium thioglycolate was omitted). As a control, the inoculum wasadded to liquid aliquot without silver and incubated at 37 C as above.Next day, the control sample was plated on agar plates and againincubated at 37 C for 24 h to 48 h to let the bacterial colonies growand become visible. From the bacterial count of the control sample andthe count of surviving colonies of the test samples, the log reduction,a quantitative measure of the antimicrobial activity was calculated.

Sterilization

A majority of the hydrogel prototypes of the present disclosure weretested for their ability to withstand elevated temperatures simply bysubjecting them to one steam sterilization cycle. The test samplesexperienced temperature rises from 20 C to 122 C over 15 min, followedby constant temperature of 122 C for 15 min and finally a cool down from122 C to ˜40 C over 3 h. Thus, the test samples experienced nearly 3.5 hof hostile temperature condition.

Physical Properties

For a thixotropic material, viscosity is not a good measure of thecharacteristic yield stress that is overcome before it begins to flow.The yield stress was determined on a rheometer such as a cone and plateor parallel plate type by conducting strain and frequency sweeps. Thistechnique is known to those ordinarily skilled in the art of physicalproperties characterization. For measuring traditional viscosity, aconcentric cylinder viscometer (with variable spindle set) such asBrookfield viscometer (Model LVDVE115) was employed.

DEFINITIONS

In the following paragraph, various terms are defined in the context ofthe present disclosure;

“Low level” of silver means a silver content <1000 ppm by weight

“Moderate level” of silver means a silver content between 1000 ppm and2000 ppm by weight

“High level” of silver means a silver content >2000 ppm by weight

“Sunlight resistant” or “light stable” or “inert to light” is defined ashaving no visible sign of discoloration (color change following exposurethat will add shade of black, brown, yellow or purple) following one oftwo exposures: (i) 30 days continuously under a 60 W incandescent tablelamp (turned on) at a distance of 12″-15″ or (ii) one hour of continuoussunlight exposure at 45N latitude.

“Steam sterilizable” or “heat stable” or “inert to heat” is defined ashaving no visible sign of discoloration (color change following exposurethat will add shade of black, brown, yellow or purple) after one steamsterilization cycle (122 C for 15 min) that includes periods of warmingup and final cool down to room temperature (<40 C).

The words “compounds” and “derivatives” in the context of silvercyanurates of the present disclosure mean the same unless the contextclearly dictates otherwise.

It should be noted that as used in this specification and the appendedclaims, the singular forms “a”, “an” and “the” include plural referentsunless the context clearly dictates otherwise.

All patents, patent applications and references included herein arespecifically incorporated by reference in their entireties.

It should be understood, of course, that the foregoing relates toexemplary embodiments of the present disclosure and that numerousmodifications or alterations may be made therein without departing fromthe spirit and the scope of the disclosure as set forth in thedisclosure.

Although the exemplary embodiments of the present disclosure areprovided herein, the present disclosure is not limited to theseembodiments. There are numerous modifications and alterations that maysuggest themselves to those skilled in the art.

The present disclosure is further illustrated by the way of the examplescontained herein, which are provided for clarity of understanding. Theexemplary embodiments should not be construed in any way as imposinglimitations upon the scope thereof. On the contrary, it is to be clearlyunderstood that resort may be had to various other embodiments,modifications, and equivalents thereof which, after reading thedescription herein, may suggest themselves to those skilled in the artwithout departing from the spirit of the present disclosure and/or thescope of the appended claims.

ILLUSTRATIVE EXAMPLES Example 1 Preparation of Mono- and Di-SodiumCyanurate

To a 50 ml PP tube, deionized water (˜23 ml) and pellets of solid sodiumhydroxide (˜0.0922 g, Sigma-Aldrich) were added. The tube was vortexedto dissolve the solids and yield a ˜0.1M solution. Next, cyanuric acidpowder (˜0.297 g) was added to the tube corresponding to 1:1 molar ratiowith respect to sodium hydroxide. The contents were vortexed and heatedin a microwave oven to dissolve the acid powder to form sodiumcyanurate. The contents were cooled to room temperature and then thetube transferred to a refrigerator. After 24 h, the tube was examinedand was found to contain fine needles of sodium cyanurate hydrate(NaH₂C₃N₃O₃.H₂O) which is consistent with a published report (see SeiferG. B., Russian Journal of Coordination Chemistry, Vol. 28, No. 5, p301-324 (2002)). The TGA scan confirmed the presence of one molecule ofwater of crystallization that was lost ˜175 C. Element anal: theor: C,21.30%; H, 2.37%; N, 24.85%; Na, 13.61%; actual: C, 21.26%; H, 2.51%; N,24.51%; Na, 13.10%.

The di-sodium cyanurate was obtained as follows. To a 400 ml glassbeaker with stir bar, cyanuric acid (2.58 g, 20 mmol), deionized water(30 ml) and sodium hydroxide solution (10 ml, 7.5M) were added and thecontents heated to ˜80 C to yield a clear solution and maintained at80-85 C for 1 h. Periodically water was added to maintain the sameliquid volume. After the heating for one hour, the liquid was cooled toroom temperature. Very small amount of solids that were present werefiltered off. A portion of the filtrate (10 ml) was transferred to asecond glass beaker (˜250 ml capacity) and heated to remove most of thewater. Solids appeared when ˜1-2 ml liquid was left in the beaker thatwas then cooled to room temperature. To the solids, aqueous ethanol (40ml, 50% v/v) were added and the solids were recovered and dried (˜0.42g). The ethanolic filtrate was left in the refrigerator (˜4 C) overnightto crystallize more needle like solids that were filtered and dried(˜0.33 g). The TGA scan of di-sodium cyanurate showed it was present asmonohydrate (Na₂HC₃N₃O₃.H₂O) with water loss at ˜110 C. Element anal:theor: C, 18.84%; H, 1.57%; N, 21.98%; Na, 24.08%; actual: C, 18.89%; H,1.58%; N, 21.66%; Na, 23.60%.

Example 2 Synthesis of Ammonium Cyanurate Compound

In a 100 ml glass Erlenmeyer flask, cyanuric acid (0.645 g, 5 mmol,Sigma Aldrich) and deionized water (10 ml) were added. Ammoniumhydroxide solution (0.4 ml, 14.8M) was pipetted into the acid watermixture. The contents were heated in a microwave oven in several 15 sdurations to obtain a clear solution. Within minutes when the hotsolution was left to cool down, fine crystals began to appear. The flaskwas covered with Saran® wrap film and left ˜4 C in a refrigeratorovernight. Next day, the crystals were recovered by filtration, washedand dried at 45 C for several hours (0.47 g, yield ˜64%). The TGA scanshowed a loss of ammonia consistent (theor: 11.64%, actual: 11.96%) withpublished work. No water of hydration was associated with the compound.

Example 3 Preparation of Gel Containing Mixed Silver Cyanurate asAntimicrobial Active Compound

The preparation of amorphous aqueous gel with mixed silver cyanuratederivative was carried out as follows. According to the published paper(by Seifer G. B. and Tarasova, Z. A., Zh. Neorg. Khim., Vol. 34, pp.1840-43, 1989), addition of soluble silver salt solution to monosodiumcyanurate solution in 1:1 mole ratio results in a variable compositionwhich was broadly termed as mixed silver cyanurate.

The first step was to prepare stock monosodium cyanurate solution. To a50 ml polypropylene conical bottom tube (BD Falcon), monosodiumcyanurate hydrate (˜0.169 g, 1 mmol) was added followed by ˜10 mldeionized water. The tube was capped and the contents briefly vortexed.Then, the tube contents were heated carefully in a microwave oven(Panasonic 1200 W) while taking care that the contents did not boilover. The hot contents were vortexed to dissolve the salt to yield aclear solution comprising monosodium cyanurate (0.1M).

In a 100 ml polypropylene plastic cup, weighed quantities of LaponiteXLG (˜0.4 g, Southern Clay Products, Gonzales, Tex.), Hydroxyethylcellulose (˜0.1 g, Lotioncrafter Inc.) and Glycerol (˜2.0 g,Lotioncrafter Inc.) were added and hand-mixed with a SS spatula to forma paste. In a second similar cup, deionized water (˜15.5 ml) was heatedto near boiling in a microwave oven. The hot water was poured into thefirst cup. Immediately thereafter, the paste was hand-mixed vigorouslywith a spatula to obtain a viscous clear gel that was left to cool toroom temperature (˜15 min).

Next, ˜1.0 ml of the monosodium cyanurate solution made above (and keptwarm at ˜40 C to prevent salt from dropping out of solution) was addedto the viscous gel and blended in to uniformity. Finally, under lowlighting conditions ˜1.0 ml aqueous silver nitrate solution (0.1M) wasadded to the gel in three aliquots; each time mixing in the aliquotbefore adding the subsequent portion. The gel became more opaque witheach aliquot addition but was pleasing and smooth to feel.

A small portion (˜3 to 4 g) was transferred to a dram vial and capped.The vial was left ˜12″ from the incandescent lamp (60 W) andcontinuously exposed to the light for 24 h. Another similar portion wastransferred to a second dram vial. The vial was capped and placed in anoven set to ˜55 C for a thermal stress test. The remaining gel wastransferred to a 50 ml PP tube and kept protected from light at roomtemperature.

After 24 h, the light exposed gel sample was examined and showed nodiscoloration. It looked identical to the gel sample kept protected fromthe light. The thermally stressed gel sample when examined after 2 weeksat 55 C showed a trace of brown color compared to gel maintained at roomtemperature. The combined results suggested that the amorphous gel madepossessed excellent resistance to discoloration induced by light andheat. In this way, the antimicrobial compound is included within anaqueous clear gel, and the aqueous clear gel with the silver cyanurateactive agent is resistant to discoloration via light. Thereby, the cleargel maintains a transparent color in response to light exposure.

Example 4 Preparation of Gel with Mixed Silver Cyanurate OmittingHydroxyethyl Cellulose as Thickener

To a 100 ml glass beaker ˜15.2 ml of deionized water were added followedby slow addition of ˜0.8 g Laponite XLG powder to the water understirring. Over the next 30 minutes, the stirred contents transformed toa clear gel. The beaker was tared and glycerol (˜2.0 g) was added whichcaused the viscosity of the gel to briefly increase. The glycerol wasstirred into the gel which did not seem to affect its clarity. Next,˜1.0 ml of warm 0.1M monosodium cyanurate solution was added and mixedinto the gel to uniformity. The gel had gained a shade of opacity thoughit remained mostly clear. Finally, as before ˜1.0 ml silver nitratesolution (0.1M) was added in roughly 3 equal aliquots to the gel. Uponcompletion of silver salt solution, we noticed slight increase in theopacity though it was much less than when hydroxyethyl cellulose wasused as thickener in the example above. Over next few days we observedthat the opacity decreased making the gel look practically clear.Though, the gel was thixotropic, it was practically transparent, smoothto feel and readily spreadable. The pH of the gel was ˜7.

Despite its clarity, the gel sample showed no discoloration after 24 hcontinuous light exposure and in appearance was similar to the gelsample protected from light. This characteristic of the gel sample isquite remarkable considering most silver salt containing gels discolorupon continuous light exposure.

When tested for antimicrobial activity against Staphylococcus aureusATCC 25923 and Pseudomonas aeruginosa ATCC 27853 in a zone of inhibitionassay, the gel sample showed clear zones confirming its activity. Thetheoretical silver content of the gel was ˜540 ppm so its antimicrobialactivity was expected. In a daily serial transfer ZOI assay, the gelsample sustained strong antimicrobial activity for 3 days against a grampositive bacteria (S. aureus ATCC 6538), a gram negative bacteria (P.aeruginosa ATCC 9027) and a yeast (C. Albicans ATCC 10231).

The sample gels thixotropic characteristic of yield stress wasdetermined using a rheometer (Rheometrics Scientific RFS Model II with25 mm parallel plates) from the strain and frequency sweeps. Themeasured value of yield stress at ˜25 C was 249 Pa.

Example 5 Comparative Example—Preparation of Gel Containing SilverSaccharinate by Method A

A comparative gel sample was prepared similar to the gel in Example 4(keeping its theoretical silver content substantially the same) exceptsilver saccharinate was the active compound. Its resistance todiscoloration by light was compared with the gel in Example 4.

The following ingredients were used: Glycerol (2.00 g), Laponite XLG(0.80 g), Saccharin (0.027 g, FW 183.2, corresponding to a slight excessover stoichiometry with respect to AgNO₃), silver nitrate (1.0 ml, 0.1M)and deionized (DI) water (16.2 g).

In a 100 ml glass beaker Laponite was dissolved in DI water to obtainclear gel as before. In a 15 ml PP tube, saccharin powder wastransferred and then glycerol was added. The tube was heated in amicrowave oven to dissolve saccharin. The resulting saccharin solutionwas dripped into the clear gel and blended into uniformity. Finally,silver nitrate solution wad added and blended into a slightly hazy gel.

A glass dram vial with a portion of the prepared gel exposed to tablelamp light from a distance of 12″ discolored to brown yellow color after16 h. The gel sample protected from light also substantially completelydiscolored to brown yellow color indicating poor resistance to lightinduced discoloration (photo-reduction).

Example 6 Comparative Example—Preparation of Silver SaccharinateContaining Gel by Method B

The gel in this example was prepared to compare its discolorationresistance with the gel in Example 4 (keeping the silver content same),but it was prepared by a different method than described in Example 5. Adifferent method was attempted to examine if it had any effect on thediscoloration resistance.

The gel was prepared in a 100 ml PP cup in a manner similar to thatdescribed in Example 4 except instead of monosodium cyanurate solution,˜1.0 ml of 0.1M sodium saccharinate solution was used.

A portion of gel in a glass dram vial discolored after 1 h upon lightexposure and in that same time period the light protected sample alsodiscolored. Both samples turned brown yellow indicating photo-reductionto silver nanoparticles. Thus, modifying the gel making procedure didnot increase the discoloration resistance.

Example 7 Comparative Example—Preparation of Silver Chloride ContainingGel

The gel was prepared as comparative example to examine if the LaponiteXLG based gel offered an environment to silver chloride as activecompound to enhance its light discoloration resistance by possiblyhelping form small crystals of chloride salt which are known to be lightinsensitive.

The gel was made by the same procedure described for the gel in Example4 except instead of monosodium cyanurate solution, 0.2M sodium chloridesolution was employed. The resulting gel was somewhat less viscous thanthe gel in Example 4 and more opaque white but smooth and pleasantlooking. The pH of the gel was ˜7.

However, upon light exposure it discolored within 2 h to a purplishblack color though the sample protected from light remained unchangedfrom its original opaque white.

Example 8 Comparative Example—Preparation of Silver Phosphate ContainingGel

The gel was prepared as in the Example 4 except instead of sodiumcyanurate solution, mono sodium phosphate solution (0.1M) was used. Therationale was to see if a silver salt of tri-functional acid such asphosphoric acid (cyanuric acid is also a tri-functional acid) affordsgreater light discoloration resistance in the gel environment.

But, the results of the light exposed gel sample showed that the samplediscolored to grey black color after 2 h. The light protected samplegave a hint of discoloration (the bright yellow shade of the gel samplehad just turned a bit dull). Overall the gel imparted a faint yellowopacity and was smooth to feel. The pH of the gel sample was between 7and 8.

Example 9 Preparation of Mixed Silver Cyanurate Containing Gel with aDifferent Formula

The following ingredients were used to prepare ˜20 g gel. Theingredients and their amounts in parentheses are listed here: Glycerol(2.00 g), Laponite XLG (0.40 g), Sodium carboxymethyl cellulose (0.1 g,Ashland Chemical Natrasol® Grade 9H4F), monosodium cyanurate solution(1.0 ml, 0.1M), silver nitrate (1.0 ml, 0.1M) and deionized (DI) water(15.5 g).

In a 100 PP cup, the solids above were blended with glycerol. DI waterwas heated to 80 C in another cup and poured into the slurry andhand-mixed as the contents cooled to room temperature. Then monosodiumcyanurate solution was added and again hand-mixed in. Finally, silvernitrate solution was added to obtain a smooth slightly opaque whiteviscous gel with silver content ˜540 ppm.

Both samples of the gel (light exposed for 24 h and the light protected)showed no discernable difference suggesting no influence of light on thesamples and both remained substantially unchanged from the time it wasmade, showing superior light discoloration resistance. With time, thegel sample seemed to show increased opacity but no discoloration. For asilver content of >500 ppm in an aqueous gel and not having any hint ofdiscoloration by light is quite extraordinary.

Example 10 Gel Sample Containing Mixed Silver Cyanurate with DifferentProportions of Thickeners

A gel sample with silver content ˜540 ppm was prepared by the sameprocedure disclosed in the Example 9 except the following amounts wereused: Glycerol (2.00 g), Laponite XLG (0.32 g), Sodium carboxymethylcellulose (0.08 g, Ashland Chemical Natrasol® Grade 9H4F), monosodiumcyanurate solution (1.0 ml, 0.1M), silver nitrate (1.0 ml, 0.1M) anddeionized (DI) water (15.6 g).

The gel was opaque white but smooth and could be spread easily. Bothsamples (24 h light exposed and light protected) showed no sign ofdiscoloration when examined nearly a month later demonstrating superiorlight discoloration resistance.

Example 11 Gel Sample Containing Mixed Silver Cyanurate with SilverContent ˜3000 ppm

Using the following ingredients and their listed proportions, the gelwas made as follows: Glycerol (2.00 g), Laponite XLG (0.80 g),monosodium cyanurate solution (5.0 ml, 0.1M), silver nitrate (5.0 ml,0.1M) and deionized (DI) water (5.3 g).

In a 100 ml PP cup, glycerol and Laponite were mixed into a paste. In a2^(nd) cup, DI water and warm monosodium cyanurate solution were mixedand poured all at once into the paste. The contents began to thicken toa gel and were hand-mixed to uniformity with a spatula. Next, silvernitrate solution was added, 0.5 ml aliquots at a time until alladditions were made (Note in this test, the lab lights were turned offduring silver nitrate addition step as a precaution). With each aliquotof silver nitrate, the opacity increased. An opaque white, smooth tofeel and pleasant gel was obtained though it was somewhat less viscousthan the gel in Example 4.

The 24 h light exposed gel sample in appearance was similar to lightprotected sample indicating superior light discoloration resistance.Even after nearly a month, there was no discoloration of the lightexposed sample though some syneresis of the gel was seen.

Example 12 Preparation of Gel Containing Mixed Silver Cyanurate withColor for Aesthetic Purposes

A gel sample was made similar to the sample in Example 4 with an addedfeature of blue color to enhance its aesthetic appeal. The blue colorwas developed by employing a copper-glutamate complex. A collateralbenefit of using copper complex was it was likely to also increase theantifungal activity of the gel via synergistic effect though withtargeted silver content of ˜540 ppm it was already lethal to fungi.

The copper-glutamate complex solution was prepared prior to the gelformulation step. Briefly, mono sodium glutamate (˜0.169 g, Ajinomotobrand obtained from a local ethnic food store) was dissolved in 10 mldeionized water. In a second PP tube, ˜0.249 g copper sulfatepentahydrate was dissolved in 10 ml DI water to obtain a bluishsolution. The two solutions were combined to yield a clear navy blueshade solution that was maintained at room temperature until ready foruse.

The following ingredients were blended in by the procedure in Example 4in the proportions indicated: Glycerol (2.00 g), Laponite XLG (0.80 g),monosodium cyanurate solution (1.0 ml, 0.1M), silver nitrate (1.0 ml,0.1M) and deionized (DI) water (14.8 g) and copper-glutamate complexsolution (0.4 ml).

As before the silver cyanurate containing gel was prepared. In the finalstep, copper-glutamate complex solution was added that imparted a faintice blue pleasing color to the gel. Qualitatively, the gel appearedsimilar to previously made gel samples containing Laponite XLG and mixedsilver cyanurate with respect to viscosity but appeared to be moretransparent. Over time it appeared that the gel sample had turned evenmore transparent. The pH of the gel was measured ˜7.

The 24 h light exposed gel samples were unaffected by continuous lightexposure and appeared substantially identical to the light protectedsample suggesting no deleterious effect of copper-glutamate complex. Thelight protected gel sample, in a daily serial transfer ZOI assay,sustained strong antimicrobial activity for 3 days against a grampositive bacteria (S. aureus ATCC 6538), a gram negative bacteria (P.aeruginosa ATCC 9027) and a yeast (C. Albicans ATCC 10231).

The measured value of yield stress (at ˜25 C) of the gel sample was 251Pa.

Example 13 Effect of Sunlight on the Gel Samples Applied Topically tothe Skin of a Human Subject

The light exposed gel samples from Example 4 and the light protected gelsample from Example 12 were applied (approx. 100 mg each) topically asthin layer on the backside of the palm of hand of a human subject andexposed directly to afternoon sunlight continuously for 15 min. and thenthe applied area was examined. No staining of the skin area was seenwhich was quite remarkable. The subject also did not experienceirritation consistent with the neutrality of the gel pH, which wasaround ˜7.

Example 14 Preparation of Sodium Silver Cyanurate Ligand ComplexContaining Gel in Large Quantity and with Excess of Monosodium Cyanurate

By pro-rating the quantities of ingredients in the Example 12, the gelon 1000 g scale was prepared with one exception. That is the monosodiumcyanurate solution amount was 20% excess over 1:1 mole ratio ofcyanurate and silver nitrate with resulting reduction in the amount ofDI water. Due to the use of an excess of cyanurate anion over silvercation, the ratio >1 was maintained. Under such situation, mixed silvercyanurate was formed. A lab stirrer/2 liter container assembly was usedto accommodate the larger proportions. The finished gel wassubstantially identical to that made in the Example 12 with silvercontent ˜540 ppm. The difference being an increased opacity compared tothe gel in Example 12.

Example 15 Preparation of Mixed Silver Cyanurate and Copper-GlutamateContaining Gels with Varying Silver Contents

Two gel samples were prepared in a manner identical to that in Example12 except the amounts of monosodium cyanurate, silver nitrate andcopper-glutamate complex solutions were altered to reflect the desiredsilver and copper contents of the finished gel samples.

One gel had theoretical target values of silver and copper as ˜110 ppmand ˜30 ppm, respectively. The second gel sample had silver and coppertarget values of ˜220 ppm and ˜60 ppm, respectively.

In appearance, these gel samples were practically transparent, verypleasing to the eyes, smooth and readily spreadable. None of the twowere adversely affected by ambient office light when examined afterseveral days on the bench, thus affirming their superior lightdiscoloration resistance.

These gel samples with lower content of silver and copper are moresuitable for treating minor cuts and wounds in the OTC market segment.

Example 16 Preparation of Alginate Fiber Based Non-Woven Dressing withMixed Silver Cyanurate

The previous examples demonstrated the superior light discolorationresistance of mixed silver cyanurate containing gels. To examine if thesame resistance was extendable to other substrates, an alginate basednon-woven dressing was treated to render it antimicrobial with mixedsilver cyanurate.

In a 15 ml PP tube, ˜11.6 ml 95% ethanol was pipetted followed by ˜0.2ml of warm clear 0.1M monosodium cyanurate and ˜0.2 ml 0.1M silvernitrate solutions. This resulted in a fine mixed silver cyanuratesuspension that was vortexed several times to keep the fine solidsevenly dispersed.

In separate petri dishes (4″ dia, BD Falcon), two ˜2″×2″ pieces ofalginate non-woven dressing (from a source in PRC) were dripped withequal portions of the mixed silver cyanurate suspension in ethanol toevenly soak them. Each piece was drained off excess ethanol by holdingit at one corner with a tweezer and the pieces were transferred to anylon mesh and left in an oven to dry for 1 h. One dry sample was leftunder the table lamp for 24 h continuous light exposure and the otherpiece was protected from light.

After 24 h, the light exposed sample had discolored to a faint purplishgray color compared to the light protected sample, but the color changewas not splotchy but uniform. Based on theory, the amount of silver inthe alginate non-woven dressing post treatment on dry basis was ˜3500ppm. A commercial non-woven alginate dressing (Maxorb® Ag) exhibitedcomparable levels of light induced discoloration. Considering that theamount of silver is so high the limited discoloration suggested themixed silver cyanurate possesses intrinsic light discolorationresistance uncommon to anti-microbial active silver compounds.

Example 17 Preparation of Mixed Silver Cyanurate Gel with PropyleneGlycol as Humectant

The gel was prepared in a manner identical to that in the Example 4except propylene glycol was used in place of glycerol as humectant. Theresulting gel appeared to be less viscous than the gel from Example 3.The gel was practically clear, smooth and readily spreadable. The gel pHwas about 7.

Upon 24 h light exposure under a table lamp, the gel sample showed nosign of discoloration consistent with previous observation with the gelin Example 4. The result suggested that propylene glycol was alsosuitable as humectant and did not adversely affect the lightdiscoloration resistance.

Example 18 Preparation of Antimicrobial Hydrogel with Mixed SilverCyanurate on 3 Kilo Scale

This example demonstrates that the gel preparation is scalable. To alarge polypropylene beaker (5 liter capacity) 2330 ml of deionized waterwas added. A high shear mixer (3″ diameter Cowles blade, IKA) wasimmersed into the water so that the blade was about 1″ above the bottom.At a stirrer speed of 800 rpm, 120 g Laponite XLG (Rockwood AdditivesCompany, South Gonzales, Tex.) was introduced quickly into the vortex ofthe stirred water through a paper cone to minimize dust cloud. The speedwas increased to ˜1100 rpm as the clay mineral dispersed and the mixturethickened. After 20 minutes stirring, a clear thixotropic gel wasobtained. Under continued stirring, Glycerol USP (300 g, LotioncrafterInc. WA) was added to the gel that briefly thinned but the viscosity wasrestored within 10 minutes. A warm solution of monosodium cyanurate (150ml, 0.1M) was dripped slowly into the thick gel and mixed in for 5minutes. This caused further thickening of the gel that was counteractedby raising the stirrer speed to ˜1600 rpm. Finally, under continuedstirring and dim light conditions, freshly prepared silver nitratesolution (75 ml, 0.2M) was dripped into the gel followed by rinse water(75 ml) used to rinse beakers containing salt solutions. The silver saltsolution was thoroughly mixed in for 30 minutes to ensure completion ofreaction such as, for example, cation exchange. Due to viscositybuildup, the stirrer was stopped several times and the semi-solids werehand-mixed with an 8″ long flat SS blade spatula to yield a smooth clearto hazy gel. After waiting overnight to ensure there was no gelsyneresis, a portion of the gel was set aside for testing and the restwas packaged in 1.5 oz polypropylene tubes. The theoretical silvercontent of the gel was ˜540 ppm.

Steam sterilization of the prepared gel was carried out as follows.About ˜20 g gel was transferred to a 50 ml PP tube (BD Falcon) that wasloosely capped and placed in an autoclave and subjected to steamsterilization (121-122 C for 15 minutes). The gel containing PP tubethat now was slightly warped was removed from the cooled autoclave andexamined. Except for a slight increase in its opacity, no discolorationof the gel mass was seen. In a subsequent test, the steam sterilized gelsample was found to be antimicrobial against Staphylococcus aureus ATCC6538 for 3 days in a serial transfer ZOI assay. Such maintenance ofantimicrobial activity in silver containing gel that experiencedelevated temperatures is quite remarkable.

Example 19 Broad Spectrum Antimicrobial Activity Testing of Gel withSilver Content ˜540 ppm

The broad spectrum antimicrobial activity of the gel of Example 18 wastested against 13 different microorganisms in a ZOI assay. Briefly,streaks of bacterial cultures were laid on agar plates and perpendicularto the streaks, gel sample was applied over as continuous string. Asnegative control, gel without silver was used. A silver containing gelsold in the market, Normlgel®Ag (Ag˜1100 ppm) served as positivecontrol.

The test results are tabulated below.

Table for Ex. 19: Broad spectrum test results on gel with mixed silvercyanurate using ZOI Assay Zone width (mm) Gel sample Exam- Nega- S pletive Positive No. Micro-organism 18 Control Control 1 Bacillus subtilisATCC 11774 3 0 4 2 Klebsiella pneumoniae ATCC 33495 3 0 2 3 Methicillinresistant Staphylococcus 4 0 3 aureus ATCC 33592 4 Enterococcus faeciumATCC 700221 7 0 6 5 Enterococcus faecalis ATCC 19433 3 0 2 6Enterobacter cloacae ATCC 13047 3 0 2 7 Proteus mirabilis ATCC 12453 3 03 8 Candida parapsilosis ATCC 22019 12.5 0 9 9 Serratia marcescens ATCC13880 3 0 3 10 Acinetobacter baumanni ATCC 5 0 5 BAA-1605 11 EscherichiaColi ATCC 8739 3 0 2 12 Listeria Monocytogenes ATCC 6 0 4 19115 13Streptococcus pyogenes ATCC 10 0 10 19615

Example 20 Preparation of Antimicrobial Hydrogel with Mixed SilverCyanurate on 3 Kilo Scale and Silver Content ˜470 ppm

The gel was prepared using the same equipment and procedure of Example 4except the ingredients and their amounts listed here were employed:Deionized water (2700 g), Laponite XLG (120 g), glycerol (300 g),monosodium cyanurate solution (150 ml, 0.1M) and silver nitrate solution(150 ml, 0.1M). The theoretical silver content of the gel was ˜470 ppm.

The gel was transferred to a white plastic PP pail and stored at ambienttemperature for 9 months with temperature fluctuating between ˜4 C and35 C. Each month the gel mass was examined for any changes. Other thanminor syneresis, no physical changes (viscosity, clarity, color orgrowth of micro-organisms) were observed. In packaged tubes however, nosyneresis was observed over the same duration.

Example 21 Preparation of Gel with Theoretical Silver Content ˜440 ppmand the Ratio of Cyanurate to Silver Ions 1.2

The rationale was to see if the ratio when skewed in favor of anioncyanurate increases the light discoloration resistance. Due to thecyanurate ions to silver ions ratio >1.0, the majority active compoundis mixed silver cyanurate. In a 100 ml PP beaker, deionized water (15.4ml) was pipetted and a magnetic stir bar was placed. Weighed quantity(0.8 g) of Laponite XLG clay mineral was quickly added to the stirredwater and the contents stirred for 10 minutes to hydrate the clay toyield a viscous gel. Thereafter, glycerol (2.0 g), monosodium cyanuratesolution 0.1M, 1 ml) and silver nitrate solution (0.1 M, 0.8 ml) wereadded in succession, mixing the ingredient thoroughly after eachaddition to obtain a smooth but slightly hazy and pleasant gel. A smallamount of gel (˜2.0 g) was transferred to a 15 ml PP tube that wascapped and placed in direct sunlight. When examined after 4 h when thetest was stopped, the gel remained clear with no discoloration. In afollow up test, gel (˜2.0 g) was taken in another 15 ml PP tube andsteam sterilized at 121-122 C for 15 minutes and then cooled to roomtemperature. Despite exposure to elevated temperature that could beconstrued as extreme for a silver containing aqueous composition, thegel remained unchanged, that is showed no sign of discoloration orchange in viscosity.

Example 22 Preparation of Gels with Various Silver Cyanurate Derivativeswith Theoretical Silver Content ˜540 ppm

Gels were prepared with mono-silver cyanurate and di-silver cyanurate asantimicrobial actives. Their suspensions were made by maintainingappropriate mole ratio of corresponding sodium cyanurate to silvernitrate that were then dripped into the base gel. By reversing the orderof addition, additional gels were prepared with mixed silver cyanuratecompounds obtained in situ by adding silver nitrate solution into basegel containing mono-, di-sodium cyanurate respectively in 1:1 moleratio. The base gel comprised of glycerol, Laponite XLG and deionizedwater. In finished gel formulations, the weight percent of glycerol,Laponite XLG and silver were ˜10%, ˜4% and ˜0.054% respectively.

All gel samples were either transparent or hazy but smooth, thixotropicand easy to spread. The pH of all gels was between 7 and 8 units. Thegels did not discolor even after continuous table lamp light exposurefor 1 week. When steam sterilized, of the four gel samples onlydi-silver cyanurate containing gel showed discoloration. Separately,when small gel samples in PP tubes were exposed to direct sunlight, onlythe di-silver cyanurate comprising gel showed hint of grey 1.5 hrespectively. Still, the discoloration resistance was observed to bequite strong. The remaining three gel samples showed no evidence ofdiscoloration through 2.5 h when the exposure was discontinued.

Example 23 Comparison of Performance of Gels Made with Silver CyanurateDerivatives and Various Silver Salts with Respect to Light Exposure

On 20 g scale, gel samples were prepared by mixing appropriate amountsof glycerol, Laponite XLG clay and water. Finally, silver salts wereformed in-situ by blending corresponding sodium salt solutions (0.1M)which was followed by silver nitrate solution (0.1M). Note in all gelpreparations, silver nitrate was added under dark. The finished gelscontained glycerol, clay and silver at 10%, 4% and 0.054% weight,respectively. The finished gel samples were observed for discolorationor lack thereof when made fresh, at 24 h and after either 1 week or 30days after being continuously exposed to a 60 W table lamp at a distanceof 1 to 1.5 feet. The results are summarized in the table below.

The results showed that of the 22 silver salts tested, silversulfadiazine and the two silver cyanurate derivatives in the gel did notshow discoloration at 24 h. But after 1 week exposure, the gel withsilver sulfadiazine showed a hint of discoloration. Thereby, the gelscomprising the silver compounds comprising s-triazine ring showedexceptional resistance to discoloration by light as evidenced by nocolor change after 4 weeks of light exposure.

Example 24 Preparation of Gel with Urea as Humectant

As before, urea (2.0 g), Laponite XLG (0.8 g) and deionized water (15.2ml) were mixed to obtain the base gel. To this, warm monosodiumcyanurate solution (1.0 ml, 0.1M) was added followed by silver nitratesolution (1.0 ml, 0.1M) in the dark. After blending the contentsthoroughly, a transparent to hazy gel was obtained. When exposed todirect sunlight for 3 h the gel showed no hint of discoloration which isquite remarkable. No discoloration of the gel was seen after steamsterilization.

Table for Ex. 23 Light Exposure Test Data on Gels with Different SilverCompounds Gel color after continuous light exposure Silver CompoundFresh After 24 h After 30 days Silver Chloride Purplish black after DarkPurplish Not tested 2 h black Silver Carbonate Hint of brown after Brownblack Not tested 10 min Silver Phosphate Grey black after 2 h Grey blackNot tested Silver Saccharinate Yellow brown Dark amber Not tested brownSilver Acetyl Salicylate Hint of grey Brown black Not tested SilverMono-tartrate Hazy gel Brown black Not tested Silver Mono-maleate Smoothhazy gel Brown yellow Not tested Silver Lactate Brown during Not testedNot tested preparation Silver Salicylate Brown after 0.5 h Not testedNot tested Silver Propionate Hint of brown after Brown black Not tested10 min Silver Sulfo-succinate Brown after 0.5 h Brown black Not testedSilver Benzene sulfonate Brown after 0.5 h Brown black Not tested SilverMono-succinate Trace of yellow Brown black Not tested grey SilverGluconate Light yellow after Brown black Not tested 10 min SilverSorbate Hint of yellow grey Brown black Not tested after 1.5 h SilverOleate Opaque white; Light yellow Not tested turned dull yellow brownafter 1 h Silver Glycolate Hint of brown grey Brown grey Not testedSilver Benzoate Light brown after Brown black Not tested 10 min SilverSulfadiazine Opaque white Hint of grey Slightly more grey SilverItaconate Hint of yellow Brown black Not tested brown C₃N₃H₂O₃Ag Hint ofhaze but No change, No change, nearly transparent nearly transparenttransparent C₃N₃O₃HAg₂ Hint of haze but No change, No change, nearlytransparent nearly transparent transparent Na[Ag(C₃N₃O₃H₂)₂] Hint ofhaze but No change, No change, nearly transparent nearly transparenttransparent NaAgHC₃N₃O₃ Hint of haze but No change, No change, nearlytransparent nearly transparent transparent

Example 25 Thermal Stability of Gels Containing Silver CyanurateDerivatives and Silver Sulfadiazine

Gels with mixed silver cyanurate and sodium silver cyanurate ligandcomplex were prepared by the method described in Example 4. With silversulfadiazine, the micronized powder corresponding to ˜540 ppm silver wasadded into the gel. The gel samples were exposed to elevatedtemperatures by subjecting them to steam sterilization. The theoreticalamount of silver in the gels was varied. The results are tabulatedbelow. As described herein, the aqueous clear gels with silver cyanurateare further resistant to discoloration via heat.

Table for Ex. 25: Observations of hydrogels with silver cyanuratecompounds after steam sterilization Theor. Silver Ratio of Color beforesteam Color after steam content (ppm) cyanurate/silver ions Activecompound sterilization sterilization 540 1 Mixed silver cyanurate HazyNo discoloration 2700 1 Mixed silver cyanurate Opaque white Nodiscoloration 440 1.25 Mixed silver cyanurate Hazy to clear Nodiscoloration 220 2.5 Sodium silver Hazy to clear No discolorationcyanurate ligand complex 110 5 Sodium silver Hazy to clear Nodiscoloration cyanurate ligand complex 540 NA Silver sulfadiazine Opaquewhite Discolored to brown

Example 26 Preparation of Talc Powder Impregnated with Silver CyanurateDerivative (Ag ˜1000 ppm)

This example demonstrates that antimicrobial property can be easilyimparted to an inorganic solid support matrix which then can be blendedinto variety of other solid articles such as catheters, plugs etc. orcan be made into coatings for surface application.

Unscented talc powder (2.5 g) was transferred to a plastic beaker (150ml capacity) with stir bar. Ethanol (5 ml) was poured over to wet thepowder and then deionized water (30 ml) was further added. Understirring and dark conditions, warm monosodium cyanurate solution (0.5ml, 0.1M) was added and immediately followed by silver nitrate solution(0.45 ml, 0.1M) to precipitate out a mixture of mixed sodium silvercynaurate ligand and variable composition silver cyanurate in thepresence of talc. The contents were stirred for 1 h in the dark.Thereafter, the talc suspension was centrifuged. The supernatant wasdiscarded, fresh ethanol (45 ml) was added. The contents vortexed tore-suspend the solids and then re-centrifuged. The liquid over thesolids was discarded and fresh ethanol was added and the contentsvortexed again and then filtered and the solids dried in an oven at 45 Cfor 2-3 h. A portion of silver impregnated talc powder in a petri-dishwas exposed to table lamp light continuously for 60 days. No change incolor was seen between exposed and light protected talc powder. Thesilver impregnated talc can be used to absorb excessive moisture due tosweat from the feet and also can be used for relief from and theeradication of fungi responsible for athlete's foot. In place of talc,one can also use zinc oxide powder or titanium oxide powder and avariety of other inorganic supports to produce antimicrobial powders forboth medical and industrial applications. In this way, the antimicrobialcompound may be further included within a solid substrate, wherein thesolid substrate is selected from the group consisting of talc powder,zinc oxide power, and titanium oxide powder. The antimicrobial functionmay also be imparted to articles, objects or surfaces by, for example,vacuum deposition of the silver cyanurate derivatives either singly oras a mixture. In this way, the methods are not limited to traditionalpreparation methods but also include methods like vacuum deposition.

Example 27 Preparation of Mixed Sodium Silver Cyanurate Ligand ComplexImpregnated Fibrous Substrates

In this illustrative example, we demonstrated the impregnation ofabsorbent paper (Bounty® brand) and cotton gauze (J&J) withantimicrobial silver cyanurate derivative at theoretical silver loadingof ˜1400 ppm and ˜500 ppm, respectively. As described, the antimicrobialsilver cyanurate compounds may be embedded within a wound dressing, acotton gauze, and/or an absorbent paper.

First the silver containing suspension was prepared as follows. In a 50ml PP tube, mono sodium cyanurate solution (5 ml, 0.1M) and silvernitrate solution (1 ml, 0.1M) were added in that order to produce afluffy white precipitate that was broken down by vortex mixing the tubecontents. After that, dilute ammonium hydroxide (25 ml, 0.3M) was addedand the contents vortexed again. To a 4″×4″ piece of paper, 4 ml of thesuspension was applied with a pipette and the piece was transferred to anylon mesh support and dried in an oven at 45 C for 1 h. A 2″×2″ cottongauze was weighed and then placed in a petri-dish. An aliquot of thesuspension substantially equal to its weight was applied to the cottongauze which was then transferred to another nylon mesh and dried at 45 Cfor 1 h.

The silver impregnated paper and gauze samples were cut in half; onepart was saved protected from light and the remaining was continuouslyexposed to table lamp light for a period of 45 days during which it wasmonitored for discoloration. No discoloration of paper or the gauzewhatsoever was observed. Separately, silver impregnated paper piece wasexposed to direct sunlight for 6 h without any discernablediscoloration. The exposed samples in ZOI assay were found antimicrobialand were effective against Staphylococcus aureus ATCC6538 andPseudomonas aeruginosa ATCC9027. Such robust discoloration resistanceagainst light have not been observed in the past for any silvercontaining products. When steam sterilized in foil pouches, silvercontaining samples of paper and cotton gauze were practically unchangedin appearance when compared to corresponding non-steam sterilizedsamples.

Example 28 Preparation of Silver Impregnated Water Glass Coating

This example illustrates the preparation of coating made of sodiumsilicate embedded with mixed silver cyanurate. To a 15 ml PP tube,deionized water (0.5 ml), sodium cyanurate solution (0.11 ml, 0.1M) andsilver nitrate solution (0.11 ml, 0.033M) were added in that order toyield a fluffy white precipitate of mixed silver cyanurate. The tubecontents were vortexed to produce a uniform suspension. In a second PPtube, ˜1 g of 40% aqueous sodium silicate solution (Raku Gold PotteryCo.) was transferred. To the silicate solution, all of the mixed silvercyanurate suspension was added and vortexed to uniformity to yield apearlescent viscous solution.

A wet coating of the viscous solution was formed on a clean glass slide(1″×4″, Fisher Scientific). The slide was placed in an oven at ˜80-100 Cfor 1 h to cure the coating embedded with silver. A hazy hard coatingwith several fissures (small and large) was obtained. The slide withcoating was left under the table lamp for continuous light exposure.After 60 days exposure no visible discoloration was observed. Thereafterthe coated slide was tested against Staphylococcus aureus ATCC6538 forantimicrobial activity by ZOI assay. It showed clear inhibition zonesindicating positive antimicrobial activity.

Example 29 Preparation of Gel with Silver Nitrate-Melamine Complex withSilver Content-540 ppm

In a 100 ml PP cup, the following ingredients and solutions were mixed:glycerol (2 g), Laponite XLG (0.8 g), deionized water (15.2 g), melamine(1 ml, 0.1M), silver nitrate (1 ml, 0.1M) to produce a smooth slightlyhazy thixotropic gel. Exposure to direct sunlight for 4 h did notproduce any discoloration of the gel, though steam sterilization turnedthe gel brown black. The gel was antimicrobial in ZOI assay.

Example 30 Preparation of Gel Sheet Material Containing Mixed SilverCyanurate and Sodium Alginate

As a first step, thixotropic gel (20 g) containing mixed silvercyanurate was prepared following the method of Example 21 havingtheoretical silver content 440 ppm. In the second step, in a PP cup,sodium alginate (0.2 g, Sigma Aldrich) was dissolved in 10 ml hotdeionized water and hand-mixed to a viscous semi-solid gel. To theresulting sodium alginate solution, 4 g of gel with silver was added andthoroughly mixed to uniformity. About ˜7 g of the resulting mixture waspoured into a ˜2″ dia plastic petri-dish and left to dry at roomtemperature over 2-3 days. A round flexible gel sheet piece weighing ˜2g was removed from the petri-dish; half of which was left exposed tooffice light for 120 days and the other half was sealed in a foil pouchand autoclaved. The light exposed piece did not undergo any change incolor over four months, but the autoclaved piece turned uniformly orangebrown. The gel sheet could suitably be used as antimicrobial dressing.The theoretical silver content of gel sheet was calculated ˜800 ppm.

Example 31 Impregnation of Pre-Made Gel Sheet with Monosilver CyanurateCompound

A piece of pre-made gel sheet (prepared according to the U.S. Pat. No.5,196,190) weighing ˜0.5 g was placed in a petri-dish. A solution madeby mixing silver nitrate (0.1 ml, 0.1M), deionized water (1.0 ml) anddilute ammonium hydroxide (1.0 ml, 0.3M) was spread evenly over thepiece to hydrate it for 30 min. Next, cyanuric acid solution (1.0 ml,0.3M) was spread of the same piece to soak up the acid. The piece wasleft protected from light for 1-2 h at room temperature. The resultingpiece imparted a faint opaque white color. The piece was found to beantimicrobial against Staphylococcus aureus ATCC3528 for 3 days in aserial transfer ZOI assay.

Example 32 Preparation of Gels with Various Silver Cyanurate Derivativesas Actives at Silver Content of 540 ppm

Each sample gel was prepared on 20 g scale. First, monosodium anddisodium salts of cyanuric acid were prepared as described in Example 1and then their 0.1M solutions were prepared. Next, the base gels weremade by hydrating LaponiteXLG (0.8 g) in deionized water (15.2 ml)followed by glycerol addition (2.0 g). Finally the gels with activecompound were made by adding silver nitrate solution and the cyanuratein the order described.

-   -   (a) For monosilver cyanurate active, monosodium cyanurate (1.0        ml, 0.1M) was added to silver nitrate (1.0 ml, 0.1M) in a        separate PP tube to produce a white suspension that was then        blended into the base gel uniformly to produce a smooth opaque        white gel.    -   (b) For mixed silver cyanurate active, monosodium cyanurate (1.0        ml, 0.1M) was directly added to the base gel, blended in        uniformly followed by silver nitrate (1.0 ml, 0.1M) and mixed to        uniformity to obtain a hazy to transparent gel.    -   (c) For sodium silver cyanurate ligand complex as active,        monosodium cyanurate (1.0 ml, 0.1M) was directly added to the        base gel, blended in uniformly followed by silver nitrate (0.5        ml, 0.1M) and make up deionized water and mixed to uniformity to        obtain a hazy to transparent gel.    -   (d) For di-silver cyanurate active, disodium cyanurate (0.5 ml,        0.1M), deionized water (0.5 ml) was added to silver nitrate (1.0        ml, 0.1M) in a separate PP tube to produce a white suspension        that was then blended into the base gel uniformly to produce a        smooth faint opaque white gel.    -   (e) For mixed sodium silver cyanurate salt active, disodium        cyanurate (1.0 ml, 0.1M) was directly added to the base gel,        blended in uniformly followed by silver nitrate (1.0 ml, 0.1M)        and further mixed to uniformity to obtain a hazy to transparent        gel.

Each of the gel samples was continuously exposed to table lamp light for1 week. No discoloration was observed. Separate samples under SLE testshowed no discoloration for exposure up to 2.5 h except the sample gel(d) above which discolored after 1.5 h, In ZOI assay employingStaphylococcus aureus ATCC3528 and Pseudomonas aeruginosa ATCC9027, eachgel exhibited antimicrobial activity. Furthermore, none of the gelsamples above showed discoloration after steam sterilization and thusshowed excellent thermal stability.

Example 33 Thermal Stability of Suspensions of Various Silver CyanurateDerivatives

Because of the strong oxidizing nature of silver ions, they tend tophoto-reduce very rapidly to elemental silver (and imparting grey, brownor black color), more so in aqueous environments. Hence, silvercontaining products that contain water are very susceptible todiscoloration both by heat and light. Whether the silver cyanuratesderivatives in aqueous environments behave, that is discolor like anoverwhelming majority of silver compounds, was investigated in thistest. In addition, the steam sterilized aqueous compositions comprisingsilver cyanurate derivatives were further examined to see if theyretained their antimicrobial properties. For the test, each suspension(with specific cyanurate derivative) was prepared under dark light inseparate PP tubes (15 ml, BD Falcon) as follows:

-   -   (i) Added monosodium cyanurate (1.0 ml, 0.1M) to silver nitrate        (1.0 ml, 0.1M) and vortexed for 1-2 minutes for mono-silver        cyanurate derivative    -   (ii) Added silver nitrate (1.0 ml, 0.1M) to monosodium cyanurate        (1.0 ml, 0.1M) and vortexed for 1-2 minutes for mixed silver        cyanurate derivative    -   (iii) Added disodium cyanurate (0.5 ml, 0.1M), deionized water        (0.5 ml) to silver nitrate (1.0 ml, 0.1 M) and vortexed for 1-2        minutes for di-silver cyanurate derivative    -   (iv) Added silver nitrate (1.0 ml, 0.1M) to disodium cyanurate        (1.0 ml, 0.1M) and vortexed for 1-2 minutes for mixed sodium        silver cyanurate derivative

The PP tubes with sterilized suspensions were observed after coolingthem to room temperature. None of them showed discoloration. Thesuspension (iii) had a hint of cream color but was consideredacceptable. Paper discs (dipped in the suspension and dried) subjectedto ZOI assay against Staphylococcus aureus ATCC6538 showed clear zonesof inhibition affirming the presence of antimicrobial activity. Thus,the silver cyanurate derivatives in water retain their antimicrobialactivity despite exposure to high temperatures.

In a follow up experiment, the steps (iii) and (iv) were scaled up by afactor of ten. The resulting solids, disilver cyanurate and sodiumsilver cyanurate were recovered as white solids after discarding thesupernatant liquids, washing with warm water three times to removeunreacted reagents and drying in oven at 45 C for several hours. Therecovered solids were analyzed for elemental composition, the results ofwhich are presented below.

Ag₂C₃N₃HO₃.H₂O: theor: Ag, 59.8%; C, 9.98%; N, 11.64%; H, 0.83%; actual:Ag, 59.17%; C, 10.82%; N, 12.48%; H, 0.56%.

NaAgC₃N₃HO₃.H₂O: theor: Ag, 39.1%; Na, 8.34%; C, 13.05%; N, 15.22%; H,1.09%; actual: Ag, 38.2%; Na, 7.66%; C, 12.76%; N, 14.65%. H, 1.28%.

Example 34 Discoloration Resistance of Silver Containing Suspensionswith Different Starting Ratios of Cyanurate to Silver Ions

The resistance to discoloration of aqueous suspensions due to heat wasexamined. The suspensions were derived by maintaining different startingratios of cyanurate anions to silver cations of stock solutions ofmonosodium cyanurate (0.1M) and silver nitrate (0.1M). The resultingsuspensions were divided into two. One portion was saved protected fromlight and the other portion was steam sterilized and then subjected tocontinuous table lamp light exposure for at least 4 weeks. The testdetails and results are summarized in the table below.

Table for Ex. 34: Thermal and light stability of aqueous suspensionswith different starting ratios of cyanurate and silver ions Observationpost AgNO3 Monosodium Deionized Observation light exposure of Test Ratio0.1M cyanurate water post steam steam. steril. no. (Anion/Cation) (ml)0.1M (ml) (ml) sterilization Sample 1 0.125 8 1 0 Barely Not testeddiscolored 2 0.167 6 1 0 Barely Not tested discolored 3 0.25 4 1 0Barely Not tested discolored 4 0.33 3 1 0 No Not tested discoloration 50.5 0.67 0.33 7 No No discoloration discoloration after 6 weeks 6 0.750.67 0.44 6.9 No Not tested discoloration 7 1 0.67 0.67 6.67 No Nottested discoloration 8 2 0.67 1.33 6 No Not tested discoloration 9 50.67 3.33 4 No No discoloration discoloration after 6 weeks

Example 35 Preparation of Hydrogel Composition with 0.3% HydrogenPeroxide as Active

This examples illustrates a hydrogel composition with 0.3% hydrogenperoxide that one can use to treat acne. The synthetic clay LaponiteXLG(0.8 g) was dispersed in deionized water (17 g) under stirring to yielda clear transparent gel. Glycerol (2.0 g) was hand-mixed into the geland finally 30% hydrogen peroxide (ACS grade, Fisher Scientific) waspipetted and uniformly mixed in. In another variation, the clay wasdispersed first into a solution of sodium chloride (0.02 g in 17 mldeionized water) and the remaining procedure was used as describedabove. The latter gel sample was applied by a human subject on acnepimples on the face each evening for 2-3 days. That resulted insubstantially complete clearing of the acne pimples. In addition, theblack scar on the pimple sites also reduced rapidly as the subjectcontinued to use the gel for few more days. The subject did notexperience irritation or burning sensation on the skin due to gel use.Another subject used the thixotropic gel (with silver content of 540ppm) of Example 4 on acne pimples for 2-3 days and found the pimplescleared quickly without any sensation of burning or irritation. Thesubjected also noticed the black scar due to pimples faded withcontinued use of the gel on the affected area for few more days.

In a modification of the gel formulation, both hydrogen peroxide (˜0.3%w/w) and mixed silver cyanurate (˜540 ppm) were included as actives. Thegel formulation sample was prepared as described in Example 4 with theexception that 0.2 ml deionized water left out. In its place, 0.2 ml 30%w/w hydrogen peroxide was added in the final step and hand-mixed touniformity. The gel formulation appeared the same as the gel in Example4.

Example 36 Preparation of Gel with Silver Contents of 110 ppm and 440ppm and Corresponding Ratios of Ag⁺ to Cyanurate Ions of 0.2 and 0.8Respectively

The gel was prepared to examine if the cyanurate anion maintained inexcess during the precipitation of sodium silver cyanurate ligandcomplex affected the discoloration resistance to light. Thus, the gelwas prepared following the procedure in Example 24 except glycerol wasused in place of urea and the volumes of silver nitrate and monosodiumcyanurate solutions were accordingly adjusted to reflect the desiredions ratio. The smooth slightly hazy gel (440 ppm Ag) was exposed todirect sunlight for 4.5 h without any discoloration. After 5 h, slightgreying was observed so the sunlight exposure was stopped. The greyedgel sample was returned to the lab drawer overnight and re-examined thenext day. The greying of the gel had reversed and the gel had becomeclear again. In contrast, a commercially available gel (Curad® fromMedline Industries) with a silver content of 55 ppm discolored to paleyellow and after overnight in the dark did not reverse the yellowdiscoloration. In comparison, the gel with 110 ppm Ag and cyanurate ionsfive time Ag ions, did not discolor in sunlight even after 6 h when thetest was terminated. The gel also did not discolor after steamsterilization. Further, the sunlight exposed steam sterilized gels (110ppm and 440 ppm Ag) retained their antimicrobial activity as evidentfrom clear inhibition zones from the ZOI assay employing Staphylococcusaureus ATCC6538 and Pseudomonas ATCC9027. Thus, a slight excess ofcyanurate ions during the gel preparation increases the light induceddiscoloration resistance without affecting excellent discolorationresistance to elevated temperatures.

In a variation of the above gel composition with 440 ppm Ag, glycerolwas replaced with propylene glycol to obtain a smooth hazy gel. The geldid not discolor even after 4 h of sunlight exposure. The gel gave afirst hint of discoloration after 4.5 h sunlight exposure though thediscoloration reversed after the gel sample was kept in the darkovernight.

In another variation of the gel, the amount of silver was set at 1100ppm. The obtained gel was smooth but somewhat opaque. When exposed todirect sunlight, the first hint of discoloration appeared at 2 h whichis quite remarkable considering a commercial gel, Normlgel Ag with 1100ppm Ag discolors within minutes.

A comparative gel was made with 110 ppm Ag but using silver saccharinateas active and the ratio of saccharinate anions to silver ions of five.The gel discolored within 10 minutes when left exposed to sunlight.

In another variation, a gel with 110 ppm Ag was made but the ratio ofcyanurate ions to silver ions was kept at one. The obtained gel wasthixotropic, smooth and slightly hazy. When exposed to sunlight, thefirst appearance of discoloration was at 4.5 h. Surprisingly, thediscoloration reversed overnight when the exposed sample was left in thedark. The gel sample registered positive antimicrobial activity againstStaphylococcus aureus ATCC6538 and Pseudomonas aeruginosa ATCC9027 inZOI assay.

Example 37 Preparation of Silver Impregnated Cotton Gauze in thePresence of Sodium Cyanurate

This examples illustrates the protective effect of cyanurate anions todiscoloration induced by light. A dipping solution comprising equalvolumes of Tween 20 (15 g/l), sodium saccharinate (0.125M), silvernitrate (0.1M) and monosodium cyanurate (0.1M) was prepared by mixingthe solution in the listed order. Two gauzes (2″×2″, Medline Industries,USP Type VII) were soaked in deionized water for 15 min and squeezed toremove water and any additives from its manufacture.

In a 15 ml PP tube, ethanol (11.2 ml, 95%) was transferred and the abovedipping solution (0.8 ml) was then added. The contents were vortexed andpoured over the two gauzes placed in a petri-dish to soak the liquidsfor a few minutes. Each piece was then gently lifted from the dish todrain as much liquid out and then placed on a nylon mesh and left to dryin an oven. The finished gauze visually looked the same as an untreatedpiece.

When exposed to sunlight, a first hint of discoloration of the silverimpregnated gauze was seen at 3 h. A gauze piece made similarly butomitting the monosodium cyanurate discolored within 0.5 h.

Example 38 Preparation of Gel with 540 ppm Ag from Sodium Cyanurate andSilver Nitrate

Using the composition of gel of Example 4, 500 g gel was made. The gelwas packaged in cosmetic grade PP tubes and left at 55 C for 8 weeks forthermal aging. Each week the gel was examined for color. Nodiscoloration was observed. The aged gel samples were tested forantimicrobial property against Staphylococcus aureus ATCC6538 andPseudomonas aeruginosa ATCC9027 in ZOI assay and were found to beeffective. The gel samples were exposed to sunlight and did not showdiscoloration until after 3 h.

Example 39 Preparation of Gel with 540 ppm Ag with a Mixture ofHumectants

A gel sample (20 g) was prepared with a substantially identicalcomposition as the gel in Example 4 except the glycerol was 12.5% byweight of the total humectant content with the rest being propyleneglycol. No discoloration of the gel was observed until after 3 h ofsunlight exposure. The discoloration seemed to have reversed its courseafter the sample was left in the dark overnight.

In another variation of the gel composition, the humectant was allpropylene glycol. When exposed to gel, it did not discolor until theexposure duration reached 3.5 h.

In yet another variation of the gel composition, the gel was madewithout the use of any humectant. The resulting gel did not discolor insunlight for 6 h when the test was stopped. However, the dried form ofthe gel recovered as flaky powder (from drying the gel by removing allmoisture) discolored rapidly in sunlight. This was expected as theamount of silver based on the amount of Laponite XLG was nearly 1.8% wt.

In yet another variation of the gel composition, the total humectantamount was split equally between glycerol and propylene glycol. In amodification of the preparation method, the humectant and the clay wasmixed first and then hydrated followed by the addition of monosodiumcyanurate and silver nitrate respectively. The discoloration due tosunlight exposure was not observed until after 7 h. Thereafter, to thediscolored sample (˜2.5 g) in a PP tube, was added 30% w/w hydrogenperoxide. The contents were vortexed and left on the bench for 2 h whenthe discoloration disappeared and the gel became clear. The sample wasre-exposed to sunlight for an additional 6 h with no noticeablediscoloration. But with exposure, the opacity of the gel increased.Clearly the presence of hydrogen peroxide increased the discolorationresistance.

Example 40 Preparation of PVA Hydrogel Sheet Embedded with SilverCyanurate Derivative

To 20 g solution of 10% w/w PVA (Sigma Aldrich, 85K-124K Mol. Wt, 98+%Hydrolyzed), monosodium cyanurate (1 ml, 0.1M) and silver nitrate (1 ml,0.1M) were successively added. The silver salt solution was added in thedark. After briefly vortexing, the contents were gently centrifuged (600rpm) for 1 min to remove bubbles. The viscous mixture was poured in a 4″dia petri-dish and subjected to 3 freeze thaw cycles (20 C/−10 C/20 C).The resulting gel piece did not show the active silver compoundprecipitated out or discolored. The gel piece could be stretched withoutbreaking and could find an application as first response burn or woundcontact anti-infective dressing because of its soothing feel on skin.The theoretical silver content of the piece was ˜1000 ppm.

Example 41 Preparation of Gel with the Ratio of Cyanurate to Silver Ions0.5

The rationale for preparing a gel sample of this example was toinvestigate the effect on light discoloration resistance when thestarting ratio of cyanurate to silver ions was less than 1.0. The gelwas prepared on 20 g scale following the method of Example 4 except thevolume of monosodium cyanurate was half. The volume deficit was made upby adding deionized water. Even after 72 h continuous table lamp lightexposure, no gel discoloration was seen.

Example 42 Preparation of Gel with Laponite XLG and NaCMC as Thickenersin Equal Proportions

In this example, equal amounts of Laponite XLG and Sodium CMC were usedwith total thickener content of 2% w/w of the gel. The method of makingthe gel was similar to that of Example 3 except the clay mineral wasallowed to hydrate first and then Sodium CMC and glycerol were added. Anopaque white but pleasant gel was obtained.

The table lamp light exposure test revealed no discoloration of thesample even after 72 h continuous exposure.

Example 43 Discoloration Testing of Mixed Silver Cyanurate as DryPrecipitate

In a 15 ml PP tube, monosodium cyanurate (1.0 ml, 0.1M) and silvernitrate solutions (1.0 ml, 0.1M) were successively transferred to obtaina white precipitate. The suspension was vortexed briefly and washedthree times with 95% ethanol. After the 3^(rd) washing, the suspensionwas poured into a petri-dish and dried at 37 C for ˜4 h. Initially, thesolid layer was exposed to table lamp light for 1 h (no discoloration)and then left in the sunlight every day for a total exposure of ˜24 h.No discoloration of the solids in the petri-dish was observed. Thisresult is quite remarkable for an antimicrobial silver compound.

Example 44 Discoloration Resistance of SilverSept® Commercial Wound CareProduct

To further test discoloration resistance to light, example commercialproducts were purchased and exposed to light to enable a comparison tothe silver cyanurate derivate compounds and products thereof asdescribed herein. In a capped glass dram vial ˜2-4 g of SilverSept® gel(Lot 0J1215S) was exposed to table lamp light. At 4 h, the first hint ofyellow appeared and after 24 h, the gel imparted distinct yellow browndiscoloration. Thus, SilverSept® gel that has ˜100 ppm silverdemonstrated poor discoloration resistance to light compared to the gelprototypes based on silver cyanurate derivative compounds.

Example 45 Discoloration Resistance Testing of Gel Prototypes withSilver Content ˜540 ppm & Comparison with Commercial Silver Wound Gels

All gels were made by following the procedure of Example 4 unless statedotherwise. Appropriate quantities of humectants as indicated in thetable were added followed by monosodium cyanurate and silver nitratesolution. In the case of gel sample #3, a slightly different method wasemployed. First, the cyanuric acid in an amount of 1:1 mole ratio withrespect to silver nitrate amount (1 ml, 0.1M) was dissolved in water.Then, Laponite XLG was dispersed, silver nitrate solution was dripped inand finally the humectant propylene glycol was added. The gel samplesincluding commercial products were placed in 15 ml PP tubes and exposedto appropriate light conditions and examined for discoloration. Theresults are tabulated below.

Table for Ex. 45: Discoloration resistance test results of gelprototypes with silver content ~540 ppm. Laponite Glycerol % PropyleneGlycol Sample ID XLG % w/w w/w % w/w TLE SLE Results 1 4 7.5 2.5 24h/test Not tested No discoloration stopped 2 4 2.5 7.5 24 h/test Nottested No discoloration stopped 3 4 0 10 Not tested 0.5 h Discolorationreverses in dark overnight 4 4 10 0 Not tested 4 h No discoloration 5 410 0 Not tested 6 h Discoloration reverses in dark overnight 6 4 10 0Not tested 6 h Discoloration reverses in dark after 3 days 7 4 0 10 Nottested 6 h No discoloration 8 4 5 5 Not tested 10 h No discoloration &no delayed discoloration SilverSept Not tested <0.25 h Discolored brown,no reversal Silvasorb Not tested <0.25 h Discolored brown, reversedpartially after 3 days Normlgel Ag Not tested <0.25 h Discolored brown,no reveral TLE: Table lamp light exposure, SLE: Sunlight exposure,Duration shown is the time of onset of discoloration

Example 46 Antimicrobial Activity Testing of Suspensions of SilverCyanurate Compounds

The suspensions have potential use in preventing or inhibiting thegrowth of microorganisms that may contaminate the contact lens when heldovernight in plastic lens case. Such contamination may be due toimproper handling of lens or lens cases or lens cleaning solutions. Toeliminate the risk of harm to the lens wearer due to potential microbialcontamination, a few drops of liquid compositions comprising silver areadded to the cleaning solution containing the lens in the case and leftovernight. As one example, the silver will go to work killing anymicroorganisms that could have contaminated the case or the solution.

To simulate this real life situation, the following experiment wasperformed in the lab. In a 50 ml PP tube, 10 ml monosodium cyanuratesolution (0.001M) were added and then 10 ml silver nitrate solution(0.001M) were added dropwise (with cyanurate to silver ions ratio ˜1.0,the active compound was mixed silver cyanurate). No immediateprecipitation was seen. So the tube was left overnight in the dark tocomplete the reaction. Next day, very fine particles were observed inthe tube when the contents were vortexed. The theoretical amount ofsilver in the suspension was ˜55 ppm. Using this stock suspension, fourequal volumes of liquid compositions with silver content of ˜50, ˜25 and˜10 and ˜0 ppm with 5% TSB were prepared in four separate 5 ml PS tubes.To each tube, the same size of inoculum of Staphylococcus aureusATCC6538 was added. The tubes were incubated at 37 C overnight. The nextday (˜20 h elapsed time) the known liquid aliquots from the tubes wereplated on agar plates and incubated at 37 C for 24 to 48 h to growcolonies. The zero time inoculum strength in cfu/ml was determined byplating the inoculum and incubating the plates at 37 C over 24 to 48 h.The surviving colonies from samples with silver were counted and fromthe zero count inoculum value (˜1e5 cfu/ml), the log reductionassociated with liquids with 50, 25 and 10 ppm silver was calculated.The results showed no surviving colonies in any tube containing silverindicating greater than 99.99% reduction.

Example 47 Preparation of Antimicrobial Petroleum Based Cream withSilver Content ˜540 ppm

The antimicrobial compound may be added to a petroleum based cream. Thefirst step was to prepare the mixed silver cyanurate suspension in a 15ml PP tube by successively pipetting stock solutions of monosodiumcyanurate (0.5 ml, 0.1M) and silver nitrate (0.5 ml, 0.1M) under dimlight conditions. After waiting for 15 min.˜1 drop of Tween 20emulsifier was added to the suspension and vortexed to uniformity. To aplastic cup (˜100 ml), 10 g petroleum jelly (Vaseline® brand from alocal store) was transferred. Next, with the help of a transfer pipet,the suspension was dripped into the cup a few drops at a time. Aftereach addition of the suspension aliquot, the jelly was vigorouslyhand-mixed to blend in the silver composition. At the end, the jellyturned to an opaque white cream that was very smooth to feel. Nodiscoloration of the cream as compared to the light protected creamsample was seen after nearly 4 weeks of exposure to table lamp light.The cream was found to be active against Staphylococcus aureus ATCC6538in ZOI assay.

Example 48 Preparation of Zinc Oxide Based Antimicrobial Ointment withSilver Content ˜540 ppm

The procedure in Example 47 was repeated with Desitin® ointment in placeof petroleum jelly. The resulting cream showed some discoloration in theform of grey coating on the exposed surface after 1 week of exposure tolamp light though the light protected sample was unchanged in color fromwhen made fresh. The grey coating was on the surface with the bulk ofthe gel unchanged. Given that the ointment composition was not optimizedfor the presence of mixed silver cyanurate active compound, thediscoloration due to greying was not entirely unexpected. Nonetheless,it took over a week for visible discoloration that indicated sufficientresistance of the silver active in Desitin® ointment environment. In ZOIassay using Staphylococcus aureus ATCC6538 the ointment was found to beantimicrobial.

Example 49 (Prophetic) Mixed Silver Cyanurate Comprising AdhesiveFormulation

An adhesive formulation similar to that example disclosed in paragraph[206] US 2009/0035342 is prepared with the following modification.Instead of employing 1M solution, stock solutions of monosodiumcyanurate and silver nitrate of 0.1M are used. The rest of theproportions of all chemicals are the same. The resulting adhesive filmcomprising in situ formed mixed silver cyanurate is expected to exertantimicrobial activity and withstand discoloration by heat and light.

Example 50 (Prophetic) Preparation of Silicone Catheter Coated with aCoating Comprising Mixed Silver Cyanurate

A curable silicone coating is prepared according to the method disclosedin Example 23 US 2009/0035342 with some modifications as follows. Asuspension of mixed silver cyanurate is prepared by mixing 0.2 ml eachof 0.1M stock solutions of silver nitrate and monosodium cyanurate. Thesuspension is diluted with THF (8 ml) and poured into the 2 partsilicone coating mixture prepared in the same proportions as describedtherein. Catheter stems are coated following the described procedure andcured employing the same thermal profile to yield silicone catheterstems coated with a coating comprising silver active cyanurate compound.The coated stems are expected to be antimicrobial and to resistdiscoloration by light and heat.

Example 51 (Prophetic) Preparation of Flexible PU Foam Impregnated withMixed Silver Cyanurate

Flexible medical grade PU foam pieces (˜1″×1″ squares and ˜2 mm thick)similar to those disclosed in Example 15 of US 2009/0035342 are soakedwith a silver cyanurate suspension prepared as follows. In a 15 ml PPtube, monosodium cyanurate solution (0.25 ml, 0.1M) is added, followedby silver nitrate solution (0.25 ml, 0.1M) under dark conditions. Theresulting white suspension is vortexed briefly and transferred to a 50ml PP tube with the help of transfer pipet. The 15 ml PP tube is rinsedwith deionized water (5 ml) and content transferred to 50 ml PP tube.Finally more deionized water is added to the suspension in 50 ml PP tubefor a total volume of 20 ml. In a shallow glass dish, four foam piecesare placed and the suspension with silver compound is poured over thepieces. The pieces are allowed to absorb the suspension over 5 min. Theneach piece is separately blotted on a folded Bounty® paper to removeexcess liquid and then transferred to a nylon mesh to dry in an oven at45 C over several hours. The pieces are expected to exhibit broadspectrum antimicrobial activity and are expected to resist discolorationby heat and light.

Example 52 Preparation of Cellulose/Polyester/Rayon Gauze PadsImpregnated with Sodium Silver Cyanurate Ligand Complex

Two pieces single ply (˜1″×1″ squares) sheet of hospital grade gauzepads made of cellulose/polyester/rayon blend (J&J Red Cross® brand Lot2631A) were placed in a petri-dish. A suspension of silver cyanuratederivative compound was prepared and dripped evenly over the pieces withthe help of a transfer pipet (˜0.5 ml/piece). The suspension was made byadding silver nitrate solution (2 ml, 0.02M) to monosodium cyanuratesolution (2 ml, 0.1M) and thoroughly vortexing the same. Each suspensionsoaked piece was blotted on a folded Bounty® paper and dried in an ovenat 45 C for 30 min. There was no visible difference between the silverimpregnated gauze sheet and the virgin gauze material. No discolorationof the silver impregnated piece was observed after 6 h of directsunlight exposure or steam sterilization in a foil pouch. In appearanceit looked the same as the sample piece that was protected from light andheat. The sunlight exposed piece was tested for antimicrobial activityagainst Staphylococcus aureus ATCC6538 in a ZOI assay and was found tobe strongly effective.

As described herein, the methods according to the present disclosureinclude methods for making an antimicrobial composition with a silvercyanurate active agent, where the methods comprise: combining aviscosity enhancing agent and a water based solvent to yield a viscousgel; and adding a metal cyanurate solution and a soluble silver saltsolution to the viscous gel, where the metal cyanurate solution and thesoluble silver salt solution react to form the silver cyanurate activeagent. As further described above, this reaction may occur in situ. Insome example, the methods further comprise adding a humectant to theviscous gel, where the humectant is one or more of glycerol, propyleneglycol, polypropylene glycol, urea, polyethylene glycol, and sodiumlactate. In other examples, the methods may comprise optionally adding acoloring agent to the antimicrobial composition, where the coloringagent is one of a water soluble dye, a copper-amino acid complex, andmethylene blue. In still other examples, the methods may compriseoptionally adding a skin enhancing additive to the antimicrobialcomposition, where the skin enhancing additive includes one or more ofan oil, a fragrance, a moisturizing agent, an emollient, a toning agent,and a surfactant. The methods may further comprise optionally adding abuffer to the viscous gel and adjusting a pH of the antimicrobialcomposition to a range of 6 to 8, although other pH ranges may bedesirable based on a particular product or application of the methods.In some instances, the methods may include pre-mixing the metalcyanurate solution and soluble silver salt solution prior to addition tothe viscous gel.

With regard to the reagents used in the methods, the viscosity enhancingagent that forms the viscous gel may be one or more of a synthetic claymineral that includes Laponite®, a natural clay mineral, a celluloseether selected from the group consisting of hydroxyethyl cellulose,hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxypropyl methylcellulose, and sodium carboxymethyl cellulose, polyacrylate, a naturalgum, a chemically modified natural gum, a chemically modified celluloseether with an aliphatic chains, a synthetic gum, polyvinyl alcohol,polyvinyl pyrrolidone, polyacrylamide, and a polyaminoacid that includesone of polyaspartate and polyglutamate. The metal cyanurate solution mayinclude one of sodium cyanurate, potassium cyanurate, lithium cyanurate,calcium cyanurate, magnesium cyanurate, barium cyanurate, coppercyanurate, zinc cyanurate, aluminum cyanurate, and ammonium cyanurate.The soluble silver salt solution may include one of silver nitrate,silver acetate, silver lactate, silver citrate, silver sulfate andsilver phosphate. According to the present disclosure, the silvercyanurate active agent may be one or more of AgNO₃.C₃N₃(NH₂)₃,C₃N₃(NH₂)₂NAg₂, Ammeline-AgNO₃, Ammelide-AgNO₃, Monosilver cyanurate(C₃N₃H₂O₃Ag), Disilver cyanurate (C₃N₃HO₃Ag₂), Trisilver cyanurate(C₃N₃O₃Ag₃), a sodium silver cyanurate ligand complex Na[Ag(C₃N₃H₂O₃)₂],a potassium silver cyanurate ligand complex K[Ag(C₃N₃H₂O₃)₂], a mixedsalt of NaAgC₃N₃HO₃, a mixed salt of NaAg₂C₃N₃O₃, a mixed salt ofKAgC₃N₃HO₃, a mixed salt KAg₂C₃N₃O₃, and a hydrated species thereof,wherein an amount of silver in the silver cyanurate active agent may bebetween 10 and 5500 ppm based on a weight of the antimicrobialcomposition.

The present disclosure further relates to antimicrobial compositionsbased on the silver compounds described. Thus, according to the presentdisclosure, an antimicrobial compound may comprise silver with ans-triazine ring. The antimicrobial compound may be included within anaqueous clear gel, where the aqueous clear gel with the antimicrobialcompound is resistant to discoloration via one or more of light andheat. In some examples, the antimicrobial compound is included within asolid substrate. For example, the solid substrate may be selected fromthe group consisting of a talc powder, a zinc oxide powder, a titaniumoxide powder, a bone powder, an inorganic porous support, a ceramic, ametal, an oxide, a pellet, a flexible foam, and a short fiber, althoughother possibilities may also be possible. In other examples, theantimicrobial compound is blended into a surface coating of a medicaldevice. However, the antimicrobial compound may also be embedded withinone or more of a woven and non-woven matrix. For example, the wovenmatrix may comprise one or more of cellulose, polyester, rayon andblends thereof (e.g., a wound dressing comprising the antimicrobialcompound). As another example, the non-woven matrix may comprise fibersof one or more of alginate and cellulose (e.g., a cotton gauze and anabsorbent paper that includes the antimicrobial compound). In stillother examples, the antimicrobial compound may be included in apetroleum based cream, a suspension, a solution, a bioadhesive, apolymer solution, a lotion, an emulsion, an emulgel, a salve, anointment, a sprayable liquid, a latex, a paste, an oily suspension, awater soluble polymeric films, and a water-insoluble film capable of thesustained release of antimicrobial silver.

One particular example of the present disclosure includes a cleartopical hydrogel comprising an antimicrobial silver cyanurate activeagent produced according to the methods disclosed. To enable topicalapplication, the hydrogel is a thixotropic hydrogel that may have ayield stress in a range of 0 to 1000 Pa. Moreover, the hydrogel is inertto light and heat. In this way, the hydrogel may resist discolorationdue to exposure to one or more of light and heat. The hydrogelcomprising the antimicrobial active agent may further allow a skin colorto be maintained in response to the topical application of the hydrogelto skin. As one example, an amount of silver in the hydrogel is between50 and 1000 ppm based on a weight of the hydrogel.

Methods of treatment are further possible using the hydrogel justdescribed. Thus, methods based on applying the clear topical hydrogelcomprising an antimicrobial compound with a silver cyanurate activeagent, to an individual are possible. As such, application of the gelmay reduce a risk of infection due to HIV during sexual contact, whereinthe individual is identified as an HIV uninfected individual. However,other applications are possible, and the gel may also be applied to anindividual to treat a dermal condition, where the dermal condition isone or more of an acute wound, a chronic wound, a first degree burn, asecond degree burn, a minor cut, a wound located on a mucous membrane,acne, rosacea, jock itch, and athlete's foot.

The reading of the description by those skilled in the art would bringto mind many alterations and modifications without departing from thespirit and the scope of the description. It is to be understood that theconfigurations and/or approaches described herein are exemplary innature, and that these specific embodiments or examples are not to beconsidered in a limiting sense, because numerous variations arepossible. The specific routines or methods described may represent oneor more of any number of data collection strategies. As such, variousacts illustrated may be performed in the sequence illustrated, in othersequences, in parallel, or in some cases omitted. Likewise, the order ofthe above-described processes may be changed.

It will be appreciated that the configurations and routines disclosedherein are exemplary in nature, and that these specific embodiments arenot to be considered in a limiting sense, because numerous variationsare possible. The subject matter of the present disclosure includes allnovel and nonobvious combinations and subcombinations of the varioussystems and configurations, and other features, functions, and/orproperties disclosed herein.

The following claims particularly point out certain combinations andsub-combinations regarded as novel and non-obvious. These claims mayrefer to “an” element or “a first” element or the equivalent thereof.Such claims should be understood to include incorporation of one or moresuch elements, neither requiring nor excluding two or more suchelements. Other combinations and sub-combinations of the disclosedfeatures, functions, elements, and/or properties may be claimed throughamendment of the present claims or through presentation of new claims inthis or a related application. Such claims, whether broader, narrower,equal, or different in scope to the original claims, also are regardedas included within the subject matter of the present disclosure.

1. A method of making an antimicrobial composition with a silvercyanurate active agent, the method comprising: combining a viscosityenhancing agent and a water based solvent to yield a viscous gel; andadding a metal cyanurate solution and a soluble silver salt solution tothe viscous gel, where the metal cyanurate solution and the solublesilver salt solution react to form the silver cyanurate active agent. 2.The method of claim 1, further comprising adding a humectant to theviscous gel, where the humectant is one or more of glycerol, propyleneglycol, polypropylene glycol, urea, polyethylene glycol, and sodiumlactate.
 3. The method of claim 2, further comprising optionally addinga coloring agent to the antimicrobial composition, where the coloringagent is one of a water soluble dye, a copper-amino acid complex, andmethylene blue.
 4. The method of claim 2, further comprising optionallyadding a skin enhancing additive to the antimicrobial composition, wherethe skin enhancing additive includes one or more of an oil, a fragrance,a moisturizing agent, an emollient, a toning agent, and a surfactant. 5.The method of claim 4, further comprising optionally adding a buffer tothe viscous gel and adjusting a pH of the antimicrobial composition to arange of 6 to
 8. 6. The method of claim 1, further comprising pre-mixingthe metal cyanurate solution and soluble silver salt solution prior toaddition to the viscous gel.
 7. The method of claim 1, wherein theviscosity enhancing agent is one or more of a synthetic clay mineralthat includes Laponite®, a natural clay mineral, a cellulose etherselected from the group consisting of hydroxyethyl cellulose,hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxypropyl methylcellulose, and sodium carboxymethyl cellulose, polyacrylate, a naturalgum, a chemically modified natural gum, a chemically modified celluloseether with an aliphatic chains, a synthetic gum, polyvinyl alcohol,polyvinyl pyrrolidone, polyacrylamide, and a polyaminoacid that includesone of polyaspartate and polyglutamate.
 8. The method of claim 1,wherein the metal cyanurate solution includes one of sodium cyanurate,potassium cyanurate, lithium cyanurate, calcium cyanurate, magnesiumcyanurate, barium cyanurate, copper cyanurate, zinc cyanurate, aluminumcyanurate, and ammonium cyanurate.
 9. The method of claim 1, wherein thesoluble silver salt solution includes one of silver nitrate, silveracetate, silver lactate, silver citrate, silver sulfate and silverphosphate.
 10. The method of claim 1, wherein the silver cyanurateactive agent is one or more of AgNO₃.C₃N₃(NH₂)₃, C₃N₃(NH₂)₂NAg₂,Ammeline-AgNO₃, Ammelide-AgNO₃, Monosilver cyanurate (C₃N₃H₂O₃Ag),Disilver cyanurate (C₃N₃HO₃Ag₂), Trisilver cyanurate (C₃N₃O₃Ag₃), asodium silver cyanurate ligand complex Na[Ag(C₃N₃H₂O₃)₂], a potassiumsilver cyanurate ligand complex K[Ag(C₃N₃H₂O₃)₂], a mixed salt ofNaAgC₃N₃HO₃, a mixed salt of NaAg₂C₃N₃O₃, a mixed salt of KAgC₃N₃HO₃, amixed salt KAg₂C₃N₃O₃, and a hydrated species thereof, and wherein anamount of silver is between 10 and 5500 ppm based on a weight of theantimicrobial composition.
 11. A antimicrobial compound, comprising:silver with an s-triazine ring.
 12. The antimicrobial compound of claim11, where the antimicrobial compound is included within an aqueous cleargel, and the aqueous clear gel with the antimicrobial compound isresistant to discoloration via one or more of light and heat.
 13. Theantimicrobial compound of claim 12, where the antimicrobial compound isincluded within a solid substrate.
 14. The antimicrobial compound ofclaim 13, wherein the solid substrate is selected from the groupconsisting of a talc powder, a zinc oxide powder, a titanium oxidepowder, a bone powder, an inorganic porous support, a ceramic, a metal,an oxide, a pellet, a flexible foam, and a short fiber.
 15. Theantimicrobial compound of claim 11, where the antimicrobial compound isone of blended into a surface coating of a medical device and embeddedwithin one or more of a woven and non-woven matrix.
 16. Theantimicrobial compound of claim 15, wherein the woven matrix comprisesone or more of cellulose, polyester, rayon and blends thereof, andwherein the non-woven matrix comprises fibers of one or more of alginateand cellulose.
 17. The antimicrobial compound of claim 11, where theantimicrobial compound is included in a petroleum based cream, asuspension, a solution, a bioadhesive, a polymer solution, a lotion, anemulsion, an emulgel, a salve, an ointment, a sprayable liquid, a latex,a paste, an oily suspension, a water soluble polymeric films, and awater-insoluble film capable of the sustained release of antimicrobialsilver.
 18. A clear topical hydrogel, comprising: an antimicrobialsilver cyanurate active agent.
 19. The clear topical hydrogel of claim18, where the hydrogel is a thixotropic hydrogel that has a yield stressin a range of 0 to 1000 Pa.
 20. The clear topical hydrogel of claim 19,where the hydrogel is inert to light and heat.
 21. The clear topicalhydrogel of claim 20, where a skin color is maintained in response to atopical application of the hydrogel to the skin.
 22. The clear topicalhydrogel of claim 21, where an amount of silver in the hydrogel isbetween 50 and 1000 ppm based on a weight of the hydrogel.
 23. A method,comprising: applying a clear topical hydrogel, the gel comprising anantimicrobial compound with a silver cyanurate active agent, to anindividual.
 24. The method of claim 23, where the gel reduces a risk ofinfection due to HIV during sexual contact, and wherein the individualis identified as an HIV uninfected individual.
 25. The method of claim23, where the gel is applied to an individual to treat a dermalcondition, where the dermal condition is one or more of an acute wound,a chronic wound, a first degree burn, a second degree burn, a minor cut,a wound located on a mucous membrane, acne, rosacea, jock itch, andathlete's foot.